Thanks ... I'll reach out to him.
On 8/10/21, 1:43 PM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
There was a seminar from the MADSSCi group today and one of the speakers
was from Cytiva and spoke on magbeads for DNA. He said he could talk to
anyone that had questions about implementing magnetic beads into their
workflow for antibody purification. His contact info and Cytiva's product
is below.
<email obscured>
https://www.cytivalifesciences.com/en/us/shop/protein-analysis/protein-sample-preparation/protein-enrichment/protein-g-mag-sepharose-p-01111
On Tue, Aug 10, 2021 at 10:33 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks everyone!
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
>
> On 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of
> Bover,Laura" <email obscured> on behalf of <email obscured>>
> wrote:
>
> We collect 12 aliquots of 1 ml each and then we measure in nanodrop
> the OD. Usually the peak appears in the 4th-5th elution sample. We pool
as
> need it. Filter and eventually concentrate.
>
> Thanks!
>
> On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David
> Blum" <email obscured> on behalf of <email obscured>> wrote:
>
> WARNING: This email originated from outside of MD Anderson.
Please
> validate the sender's email address before clicking on links or
attachments
> as they may not be safe.
>
> Frances,
>
> If you don't want to use an FPLC you can use a simple syringe
with
> the
> pre-packed protein a or g columns and a luer adapter which may be
> included
> in the box with the column. Since this is a bind an elute
process
> it
> should be fine to do it this way, but you would need to determine
> the
> optimal elution volume and range since you won't have a UV
> detector.
>
> On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan
> Kettering
> Institute <email obscured>> wrote:
>
> > Thanks David.
> >
> > Frances
> >
> > ZRC 1563
> > Office 646-888-2354
> > Mobile 516-967-0656
> >
> >
> > On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of
> David Blum" <
> > <email obscured> on behalf of <email obscured>> wrote:
> >
> > Frances: I understand. This might not be the best option,
> but it
> > should
> > be good for processing multiple samples quickly without
> using the FPLC.
> > You could also batch bind with the resin you use now and
> spin down at a
> > speed that doesn't deform the beads. Good luck!
> >
> > David
> >
> > On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan
> Kettering
> > Institute <email obscured>> wrote:
> >
> > > Thanks David. The binding capacity is pretty low
relative
> to
> > standard
> > > resins and the beads are much more expensive.
> > >
> > > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic
> Beads
> > >
> > > while
> > >
> > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4
> Fast Flow
> > Media
> > > (formerly GE LifeSciences / Formerly Pharmacia ... boy am
> I old)
> > >
> > > I know I want to use the FastFlow resin (or something
> similar) ...
> > right
> > > now I am more looking for a process that does not involve
> an FPLC if
> > I can
> > > help it. I have one but the person who has used it is
> retireing and
> > this
> > > is the only thing we will use it for, so do I need it is
> the
> > question.
> > >
> > > A lab (Ivo this was Pop Council) used to grow up a 100s
of
> mls of
> > each
> > > hybridoma then line the bottles of conditioned media up
on
> a shelf,
> > > allowing the media to flow through a tube into a column
> below it and
> > pass
> > > over the a prot G resin over night by gravity. Some how
> they set it
> > up so
> > > the columns DID NOT DRY OUT. The next morning they would
> wash the
> > columns
> > > by gravitty again in series and then elute them one at a
> time.
> > Sounds like
> > > a great way to do a 5 to 20 at once, and I am sure I
could
> figure
> > this out,
> > > but before I spend time on this, was wondering if anyone
> else already
> > > tackled this or had a better mouse trap.
> > >
> > > Frances
> > >
> > > ZRC 1563
> > > Office 646-888-2354
> > > Mobile 516-967-0656
> > >
> > >
> ____________________________________________________________
> > >
> > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf
> of David
> > Blum" <
> > > <email obscured> on behalf of <email obscured>>
> wrote:
> > >
> > > Have you tried mag beads? We use them for histagged
> proteins at
> > small
> > > scale but there are protein coupled beads too.
> > >
> > >
>
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$
> > >
> > >
> > >
> > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia,
> Frances/Sloan
> > Kettering
> > > Institute <email obscured>> wrote:
> > >
> > > > Happy Friday.
> > > >
> > > > Do any of you produce and purify small amounts of
> the new MAbs
> > you
> > > make
> > > > for further characterization prior to committing to
> a few or
> > one
> > > winner?
> > > > If so, how do you do this. Currently we make 350
ml
> exhausted
> > media
> > > > (hybridoma adapted to 1% FBS prescreened for
> ultra-low levels
> > of
> > > bovine IgG
> > > > + 1 ng/ml IL6) and then purify it on a Prot G
column
> using and
> > > FPLC. We
> > > > get 1 – 8 mg, but I would like to scale this back
> and simply
> > it if
> > > possible
> > > > because we usually do not need that much.
> > > >
> > > > Thanks.
> > > >
> > > > Frances
> > > >
> > > >
> > > > Frances Weis-Garcia, Ph.D.
> > > >
> > > > Head
> > > > Bi-Institutional Antibody and Bioresource Core
> Facility
> > > > (RRID: CR 017691)
> > > >
> > > > Associate Laboratory Member
> > > > Immunology Program
> > > >
> > > > Memorial Sloan Kettering Cancer Center
> > > > 1275 York Avenue, New York, NY 10065
> > > > <
> > >
> >
>
https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLcA59hJ3$
> > > >
> > > > T 646.888.2354 C 516.967.0656
> > > >
> > > > ABCF-MSKCC website<
> > > >
> > >
> >
>
https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLfRBPtf4$
> > > > >
> > > >
> > > > Adjunct Faculty
> > > > Center for Clinical and Translational Science
> > > >
> > > > The Rockefeller University
> > > > 1230 York Avenue, New York, NY 10065
> > > > <
> > >
> >
>
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> > > >
> > > > T 212.327.7030
> > > >
> > > > ABCF-RU website<
>
https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLbgJ5Ak4$
> >
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Posts in Antibody Discussion Forum
There was a seminar from the MADSSCi group today and one of the speakers was from Cytiva and spoke on magbeads for DNA. He said he could talk to anyone that had questions about implementing magnetic beads into their workflow for antibody purification. His contact info and Cytiva's product is below. <email obscured> https://www.cytivalifesciences.com/en/us/shop/protein-analysis/protein-sample-preparation/protein-enrichment/protein-g-mag-sepharose-p-01111 On Tue, Aug 10, 2021 at 10:33 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks everyone! > > Frances > > ZRC 1563 > Office 646-888-2354 > Mobile 516-967-0656 > > > On 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of > Bover,Laura" <email obscured> on behalf of <email obscured>> > wrote: > > We collect 12 aliquots of 1 ml each and then we measure in nanodrop > the OD. Usually the peak appears in the 4th-5th elution sample. We pool as > need it. Filter and eventually concentrate. > > Thanks! > > On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David > Blum" <email obscured> on behalf of <email obscured>> wrote: > > WARNING: This email originated from outside of MD Anderson. Please > validate the sender's email address before clicking on links or attachments > as they may not be safe. > > Frances, > > If you don't want to use an FPLC you can use a simple syringe with > the > pre-packed protein a or g columns and a luer adapter which may be > included > in the box with the column. Since this is a bind an elute process > it > should be fine to do it this way, but you would need to determine > the > optimal elution volume and range since you won't have a UV > detector. > > On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan > Kettering > Institute <email obscured>> wrote: > > > Thanks David. > > > > Frances > > > > ZRC 1563 > > Office 646-888-2354 > > Mobile 516-967-0656 > > > > > > On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of > David Blum" < > > <email obscured> on behalf of <email obscured>> wrote: > > > > Frances: I understand. This might not be the best option, > but it > > should > > be good for processing multiple samples quickly without > using the FPLC. > > You could also batch bind with the resin you use now and > spin down at a > > speed that doesn't deform the beads. Good luck! > > > > David > > > > On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan > Kettering > > Institute <email obscured>> wrote: > > > > > Thanks David. The binding capacity is pretty low relative > to > > standard > > > resins and the beads are much more expensive. > > > > > > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic > Beads > > > > > > while > > > > > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 > Fast Flow > > Media > > > (formerly GE LifeSciences / Formerly Pharmacia ... boy am > I old) > > > > > > I know I want to use the FastFlow resin (or something > similar) ... > > right > > > now I am more looking for a process that does not involve > an FPLC if > > I can > > > help it. I have one but the person who has used it is > retireing and > > this > > > is the only thing we will use it for, so do I need it is > the > > question. > > > > > > A lab (Ivo this was Pop Council) used to grow up a 100s of > mls of > > each > > > hybridoma then line the bottles of conditioned media up on > a shelf, > > > allowing the media to flow through a tube into a column > below it and > > pass > > > over the a prot G resin over night by gravity. Some how > they set it > > up so > > > the columns DID NOT DRY OUT. The next morning they would > wash the > > columns > > > by gravitty again in series and then elute them one at a > time. > > Sounds like > > > a great way to do a 5 to 20 at once, and I am sure I could > figure > > this out, > > > but before I spend time on this, was wondering if anyone > else already > > > tackled this or had a better mouse trap. > > > > > > Frances > > > > > > ZRC 1563 > > > Office 646-888-2354 > > > Mobile 516-967-0656 > > > > > > > ____________________________________________________________ > > > > > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf > of David > > Blum" < > > > <email obscured> on behalf of <email obscured>> > wrote: > > > > > > Have you tried mag beads? We use them for histagged > proteins at > > small > > > scale but there are protein coupled beads too. > > > > > > > https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$ > > > > > > > > > > > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, > Frances/Sloan > > Kettering > > > Institute <email obscured>> wrote: > > > > > > > Happy Friday. > > > > > > > > Do any of you produce and purify small amounts of > the new MAbs > > you > > > make > > > > for further characterization prior to committing to > a few or > > one > > > winner? > > > > If so, how do you do this. Currently we make 350 ml > exhausted > > media > > > > (hybridoma adapted to 1% FBS prescreened for > ultra-low levels > > of > > > bovine IgG > > > > + 1 ng/ml IL6) and then purify it on a Prot G column > using and > > > FPLC. We > > > > get 1 – 8 mg, but I would like to scale this back > and simply > > it if > > > possible > > > > because we usually do not need that much. > > > > > > > > Thanks. > > > > > > > > Frances > > > > > > > > > > > > Frances Weis-Garcia, Ph.D. > > > > > > > > Head > > > > Bi-Institutional Antibody and Bioresource Core > Facility > > > > (RRID: CR 017691) > > > > > > > > Associate Laboratory Member > > > > Immunology Program > > > > > > > > Memorial Sloan Kettering Cancer Center > > > > 1275 York Avenue, New York, NY 10065 > > > > < > > > > > > https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLcA59hJ3$ > > > > > > > > T 646.888.2354 C 516.967.0656 > > > > > > > > ABCF-MSKCC website< > > > > > > > > > > https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLfRBPtf4$ > > > > > > > > > > > > > Adjunct Faculty > > > > Center for Clinical and Translational Science > > > > > > > > The Rockefeller University > > > > 1230 York Avenue, New York, NY 10065 > > > > < > > > > > > https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLZ2GrQuq$ > > > > > > > > T 212.327.7030 > > > > > > > > ABCF-RU website< > https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLbgJ5Ak4$ > > > > > > > > > > > > > > Please note that this e-mail and any files > transmitted with it > > may be > > > > privileged, confidential, and protected from > disclosure under > > > applicable > > > > law. If the reader of this message is not the > intended > > recipient, or > > > an > > > > employee or agent responsible for delivering this > message to > > the > > > intended > > > > recipient, you are hereby notified that any reading, > > dissemination, > > > > distribution, copying, or other use of this > communication or > > any of > > > its > > > > attachments is strictly prohibited. If you have > received this > > > > communication in error, please notify the sender > immediately by > > > replying to > > > > this message and deleting this message, any > attachments, and > > all > > > copies and > > > > backups from your computer. > > > > > > > > > > > > > > > ===================================================================== > > > > > > > > Please note that this e-mail and any files > transmitted > > from > > > > Memorial Sloan Kettering Cancer Center may be > privileged, > > > > confidential, > > > > and protected from disclosure under applicable > law. If the > > > reader of > > > > this message is not the intended recipient, or > an > > employee or > > > agent > > > > responsible for delivering this message to the > intended > > > recipient, > > > > you are hereby notified that any reading, > dissemination, > > > distribution, > > > > copying, or other use of this communication or > any of its > > > attachments > > > > is strictly prohibited. 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If you are not the intended > recipient, or an authorized representative of the intended recipient, any > further review, disclosure, use, dissemination, distribution, or copying of > this message or any attachment (or the information contained therein) is > strictly prohibited. If you think that you have received this e-mail > message in error, please notify the sender by return e-mail and delete all > references to it and its contents from your systems. > > ―― > View topic http://list.abrf.org/r/topic/4cAjEjoPGxhp0Rsc6ZDUR > Leave group <email obscured>?Subject=Unsubscribe > > > > ===================================================================== > > Please note that this e-mail and any files transmitted from > Memorial Sloan Kettering Cancer Center may be privileged, > confidential, > and protected from disclosure under applicable law. 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Thanks everyone!
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
We collect 12 aliquots of 1 ml each and then we measure in nanodrop the OD.
Usually the peak appears in the 4th-5th elution sample. We pool as need it.
Filter and eventually concentrate.
Thanks!
On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please
validate the sender's email address before clicking on links or attachments as
they may not be safe.
Frances,
If you don't want to use an FPLC you can use a simple syringe with the
pre-packed protein a or g columns and a luer adapter which may be
included
in the box with the column. Since this is a bind an elute process it
should be fine to do it this way, but you would need to determine the
optimal elution volume and range since you won't have a UV detector.
On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks David.
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
>
> On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of David
Blum" <
> <email obscured> on behalf of <email obscured>> wrote:
>
> Frances: I understand. This might not be the best option, but
it
> should
> be good for processing multiple samples quickly without using the
FPLC.
> You could also batch bind with the resin you use now and spin
down at a
> speed that doesn't deform the beads. Good luck!
>
> David
>
> On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan
Kettering
> Institute <email obscured>> wrote:
>
> > Thanks David. The binding capacity is pretty low relative to
> standard
> > resins and the beads are much more expensive.
> >
> > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads
> >
> > while
> >
> > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast
Flow
> Media
> > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I
old)
> >
> > I know I want to use the FastFlow resin (or something similar)
...
> right
> > now I am more looking for a process that does not involve an
FPLC if
> I can
> > help it. I have one but the person who has used it is
retireing and
> this
> > is the only thing we will use it for, so do I need it is the
> question.
> >
> > A lab (Ivo this was Pop Council) used to grow up a 100s of mls
of
> each
> > hybridoma then line the bottles of conditioned media up on a
shelf,
> > allowing the media to flow through a tube into a column below
it and
> pass
> > over the a prot G resin over night by gravity. Some how they
set it
> up so
> > the columns DID NOT DRY OUT. The next morning they would wash
the
> columns
> > by gravitty again in series and then elute them one at a time.
> Sounds like
> > a great way to do a 5 to 20 at once, and I am sure I could
figure
> this out,
> > but before I spend time on this, was wondering if anyone else
already
> > tackled this or had a better mouse trap.
> >
> > Frances
> >
> > ZRC 1563
> > Office 646-888-2354
> > Mobile 516-967-0656
> >
> > ____________________________________________________________
> >
> > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of
David
> Blum" <
> > <email obscured> on behalf of <email obscured>> wrote:
> >
> > Have you tried mag beads? We use them for histagged
proteins at
> small
> > scale but there are protein coupled beads too.
> >
> >
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$
> >
> >
> >
> > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan
> Kettering
> > Institute <email obscured>> wrote:
> >
> > > Happy Friday.
> > >
> > > Do any of you produce and purify small amounts of the new
MAbs
> you
> > make
> > > for further characterization prior to committing to a few
or
> one
> > winner?
> > > If so, how do you do this. Currently we make 350 ml
exhausted
> media
> > > (hybridoma adapted to 1% FBS prescreened for ultra-low
levels
> of
> > bovine IgG
> > > + 1 ng/ml IL6) and then purify it on a Prot G column
using and
> > FPLC. We
> > > get 1 – 8 mg, but I would like to scale this back and
simply
> it if
> > possible
> > > because we usually do not need that much.
> > >
> > > Thanks.
> > >
> > > Frances
> > >
> > >
> > > Frances Weis-Garcia, Ph.D.
> > >
> > > Head
> > > Bi-Institutional Antibody and Bioresource Core Facility
> > > (RRID: CR 017691)
> > >
> > > Associate Laboratory Member
> > > Immunology Program
> > >
> > > Memorial Sloan Kettering Cancer Center
> > > 1275 York Avenue, New York, NY 10065
> > > <
> >
>
https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLcA59hJ3$
> > >
> > > T 646.888.2354 C 516.967.0656
> > >
> > > ABCF-MSKCC website<
> > >
> >
>
https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLfRBPtf4$
> > > >
> > >
> > > Adjunct Faculty
> > > Center for Clinical and Translational Science
> > >
> > > The Rockefeller University
> > > 1230 York Avenue, New York, NY 10065
> > > <
> >
>
https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLZ2GrQuq$
> > >
> > > T 212.327.7030
> > >
> > > ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLbgJ5Ak4$
>
> > >
> > >
> > > Please note that this e-mail and any files transmitted
with it
> may be
> > > privileged, confidential, and protected from disclosure
under
> > applicable
> > > law. If the reader of this message is not the intended
> recipient, or
> > an
> > > employee or agent responsible for delivering this message
to
> the
> > intended
> > > recipient, you are hereby notified that any reading,
> dissemination,
> > > distribution, copying, or other use of this communication
or
> any of
> > its
> > > attachments is strictly prohibited. If you have received
this
> > > communication in error, please notify the sender
immediately by
> > replying to
> > > this message and deleting this message, any attachments,
and
> all
> > copies and
> > > backups from your computer.
> > >
> > >
> > >
> =====================================================================
> > >
> > > Please note that this e-mail and any files
transmitted
> from
> > > Memorial Sloan Kettering Cancer Center may be
privileged,
> > > confidential,
> > > and protected from disclosure under applicable law.
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> > reader of
> > > this message is not the intended recipient, or an
> employee or
> > agent
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> > Please note that this e-mail and any files transmitted
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> > Memorial Sloan Kettering Cancer Center may be privileged,
> > confidential,
> > and protected from disclosure under applicable law. If the
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> > this message is not the intended recipient, or an employee
or
> agent
> > responsible for delivering this message to the intended
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> =====================================================================
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> Please note that this e-mail and any files transmitted from
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reader of
> this message is not the intended recipient, or an employee or
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from your computer.
We collect 12 aliquots of 1 ml each and then we measure in nanodrop the OD.
Usually the peak appears in the 4th-5th elution sample. We pool as need it.
Filter and eventually concentrate.
Thanks!
On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Frances,
If you don't want to use an FPLC you can use a simple syringe with the
pre-packed protein a or g columns and a luer adapter which may be included
in the box with the column. Since this is a bind an elute process it
should be fine to do it this way, but you would need to determine the
optimal elution volume and range since you won't have a UV detector.
On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks David.
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
>
> On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of David Blum" <
> <email obscured> on behalf of <email obscured>> wrote:
>
> Frances: I understand. This might not be the best option, but it
> should
> be good for processing multiple samples quickly without using the
FPLC.
> You could also batch bind with the resin you use now and spin down at
a
> speed that doesn't deform the beads. Good luck!
>
> David
>
> On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
> Institute <email obscured>> wrote:
>
> > Thanks David. The binding capacity is pretty low relative to
> standard
> > resins and the beads are much more expensive.
> >
> > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads
> >
> > while
> >
> > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow
> Media
> > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old)
> >
> > I know I want to use the FastFlow resin (or something similar) ...
> right
> > now I am more looking for a process that does not involve an FPLC
if
> I can
> > help it. I have one but the person who has used it is retireing
and
> this
> > is the only thing we will use it for, so do I need it is the
> question.
> >
> > A lab (Ivo this was Pop Council) used to grow up a 100s of mls of
> each
> > hybridoma then line the bottles of conditioned media up on a shelf,
> > allowing the media to flow through a tube into a column below it
and
> pass
> > over the a prot G resin over night by gravity. Some how they set it
> up so
> > the columns DID NOT DRY OUT. The next morning they would wash the
> columns
> > by gravitty again in series and then elute them one at a time.
> Sounds like
> > a great way to do a 5 to 20 at once, and I am sure I could figure
> this out,
> > but before I spend time on this, was wondering if anyone else
already
> > tackled this or had a better mouse trap.
> >
> > Frances
> >
> > ZRC 1563
> > Office 646-888-2354
> > Mobile 516-967-0656
> >
> > ____________________________________________________________
> >
> > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David
> Blum" <
> > <email obscured> on behalf of <email obscured>> wrote:
> >
> > Have you tried mag beads? We use them for histagged proteins
at
> small
> > scale but there are protein coupled beads too.
> >
> >
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$
> >
> >
> >
> > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan
> Kettering
> > Institute <email obscured>> wrote:
> >
> > > Happy Friday.
> > >
> > > Do any of you produce and purify small amounts of the new
MAbs
> you
> > make
> > > for further characterization prior to committing to a few or
> one
> > winner?
> > > If so, how do you do this. Currently we make 350 ml
exhausted
> media
> > > (hybridoma adapted to 1% FBS prescreened for ultra-low levels
> of
> > bovine IgG
> > > + 1 ng/ml IL6) and then purify it on a Prot G column using
and
> > FPLC. We
> > > get 1 – 8 mg, but I would like to scale this back and simply
> it if
> > possible
> > > because we usually do not need that much.
> > >
> > > Thanks.
> > >
> > > Frances
> > >
> > >
> > > Frances Weis-Garcia, Ph.D.
> > >
> > > Head
> > > Bi-Institutional Antibody and Bioresource Core Facility
> > > (RRID: CR 017691)
> > >
> > > Associate Laboratory Member
> > > Immunology Program
> > >
> > > Memorial Sloan Kettering Cancer Center
> > > 1275 York Avenue, New York, NY 10065
> > > <
> >
>
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> > >
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> > >
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> > >
> >
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Frances, If you don't want to use an FPLC you can use a simple syringe with the pre-packed protein a or g columns and a luer adapter which may be included in the box with the column. Since this is a bind an elute process it should be fine to do it this way, but you would need to determine the optimal elution volume and range since you won't have a UV detector. On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks David. > > Frances > > ZRC 1563 > Office 646-888-2354 > Mobile 516-967-0656 > > > On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of David Blum" < > <email obscured> on behalf of <email obscured>> wrote: > > Frances: I understand. This might not be the best option, but it > should > be good for processing multiple samples quickly without using the FPLC. > You could also batch bind with the resin you use now and spin down at a > speed that doesn't deform the beads. Good luck! > > David > > On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering > Institute <email obscured>> wrote: > > > Thanks David. The binding capacity is pretty low relative to > standard > > resins and the beads are much more expensive. > > > > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads > > > > while > > > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow > Media > > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) > > > > I know I want to use the FastFlow resin (or something similar) ... > right > > now I am more looking for a process that does not involve an FPLC if > I can > > help it. I have one but the person who has used it is retireing and > this > > is the only thing we will use it for, so do I need it is the > question. > > > > A lab (Ivo this was Pop Council) used to grow up a 100s of mls of > each > > hybridoma then line the bottles of conditioned media up on a shelf, > > allowing the media to flow through a tube into a column below it and > pass > > over the a prot G resin over night by gravity. Some how they set it > up so > > the columns DID NOT DRY OUT. The next morning they would wash the > columns > > by gravitty again in series and then elute them one at a time. > Sounds like > > a great way to do a 5 to 20 at once, and I am sure I could figure > this out, > > but before I spend time on this, was wondering if anyone else already > > tackled this or had a better mouse trap. > > > > Frances > > > > ZRC 1563 > > Office 646-888-2354 > > Mobile 516-967-0656 > > > > ____________________________________________________________ > > > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David > Blum" < > > <email obscured> on behalf of <email obscured>> wrote: > > > > Have you tried mag beads? We use them for histagged proteins at > small > > scale but there are protein coupled beads too. > > > > https://www.thermofisher.com/order/catalog/product/88848 > > > > > > > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan > Kettering > > Institute <email obscured>> wrote: > > > > > Happy Friday. > > > > > > Do any of you produce and purify small amounts of the new MAbs > you > > make > > > for further characterization prior to committing to a few or > one > > winner? > > > If so, how do you do this. Currently we make 350 ml exhausted > media > > > (hybridoma adapted to 1% FBS prescreened for ultra-low levels > of > > bovine IgG > > > + 1 ng/ml IL6) and then purify it on a Prot G column using and > > FPLC. We > > > get 1 – 8 mg, but I would like to scale this back and simply > it if > > possible > > > because we usually do not need that much. > > > > > > Thanks. > > > > > > Frances > > > > > > > > > Frances Weis-Garcia, Ph.D. > > > > > > Head > > > Bi-Institutional Antibody and Bioresource Core Facility > > > (RRID: CR 017691) > > > > > > Associate Laboratory Member > > > Immunology Program > > > > > > Memorial Sloan Kettering Cancer Center > > > 1275 York Avenue, New York, NY 10065 > > > < > > > https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g > > > > > > T 646.888.2354 C 516.967.0656 > > > > > > ABCF-MSKCC website< > > > > > > https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility > > > > > > > > > > Adjunct Faculty > > > Center for Clinical and Translational Science > > > > > > The Rockefeller University > > > 1230 York Avenue, New York, NY 10065 > > > < > > > https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g > > > > > > T 212.327.7030 > > > > > > ABCF-RU website<https://www.rockefeller.edu/monoclonal/> > > > > > > > > > Please note that this e-mail and any files transmitted with it > may be > > > privileged, confidential, and protected from disclosure under > > applicable > > > law. If the reader of this message is not the intended > recipient, or > > an > > > employee or agent responsible for delivering this message to > the > > intended > > > recipient, you are hereby notified that any reading, > dissemination, > > > distribution, copying, or other use of this communication or > any of > > its > > > attachments is strictly prohibited. If you have received this > > > communication in error, please notify the sender immediately by > > replying to > > > this message and deleting this message, any attachments, and > all > > copies and > > > backups from your computer. > > > > > > > > > > ===================================================================== > > > > > > Please note that this e-mail and any files transmitted > from > > > Memorial Sloan Kettering Cancer Center may be privileged, > > > confidential, > > > and protected from disclosure under applicable law. If the > > reader of > > > this message is not the intended recipient, or an > employee or > > agent > > > responsible for delivering this message to the intended > > recipient, > > > you are hereby notified that any reading, dissemination, > > distribution, > > > copying, or other use of this communication or any of its > > attachments > > > is strictly prohibited. If you have received this > > communication in > > > error, please notify the sender immediately by replying > to this > > > message > > > and deleting this message, any attachments, and all > copies and > > backups > > > from your computer. > > > > > > ―― > > > View topic http://list.abrf.org/r/topic/6Md8fJOjSbIvYb2uRBONUs > > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > ―― > > View topic http://list.abrf.org/r/topic/678uk6H9gQHzJRE5YnHNKa > > Leave group <email obscured>?Subject=Unsubscribe > > > > > > > > > > *** Only open attachments or links from trusted senders. Report > > phishing to <email obscured> *** > > > > > > ===================================================================== > > > > Please note that this e-mail and any files transmitted from > > Memorial Sloan Kettering Cancer Center may be privileged, > > confidential, > > and protected from disclosure under applicable law. If the > reader of > > this message is not the intended recipient, or an employee or > agent > > responsible for delivering this message to the intended > recipient, > > you are hereby notified that any reading, dissemination, > distribution, > > copying, or other use of this communication or any of its > attachments > > is strictly prohibited. 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Thanks David.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
Frances: I understand. This might not be the best option, but it should
be good for processing multiple samples quickly without using the FPLC.
You could also batch bind with the resin you use now and spin down at a
speed that doesn't deform the beads. Good luck!
David
On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks David. The binding capacity is pretty low relative to standard
> resins and the beads are much more expensive.
>
> > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads
>
> while
>
> > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media
> (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old)
>
> I know I want to use the FastFlow resin (or something similar) ... right
> now I am more looking for a process that does not involve an FPLC if I
can
> help it. I have one but the person who has used it is retireing and this
> is the only thing we will use it for, so do I need it is the question.
>
> A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each
> hybridoma then line the bottles of conditioned media up on a shelf,
> allowing the media to flow through a tube into a column below it and pass
> over the a prot G resin over night by gravity. Some how they set it up so
> the columns DID NOT DRY OUT. The next morning they would wash the
columns
> by gravitty again in series and then elute them one at a time. Sounds
like
> a great way to do a 5 to 20 at once, and I am sure I could figure this
out,
> but before I spend time on this, was wondering if anyone else already
> tackled this or had a better mouse trap.
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
> ____________________________________________________________
>
> On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" <
> <email obscured> on behalf of <email obscured>> wrote:
>
> Have you tried mag beads? We use them for histagged proteins at
small
> scale but there are protein coupled beads too.
>
> https://www.thermofisher.com/order/catalog/product/88848
>
>
>
> On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering
> Institute <email obscured>> wrote:
>
> > Happy Friday.
> >
> > Do any of you produce and purify small amounts of the new MAbs you
> make
> > for further characterization prior to committing to a few or one
> winner?
> > If so, how do you do this. Currently we make 350 ml exhausted
media
> > (hybridoma adapted to 1% FBS prescreened for ultra-low levels of
> bovine IgG
> > + 1 ng/ml IL6) and then purify it on a Prot G column using and
> FPLC. We
> > get 1 – 8 mg, but I would like to scale this back and simply it if
> possible
> > because we usually do not need that much.
> >
> > Thanks.
> >
> > Frances
> >
> >
> > Frances Weis-Garcia, Ph.D.
> >
> > Head
> > Bi-Institutional Antibody and Bioresource Core Facility
> > (RRID: CR 017691)
> >
> > Associate Laboratory Member
> > Immunology Program
> >
> > Memorial Sloan Kettering Cancer Center
> > 1275 York Avenue, New York, NY 10065
> > <
>
https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g
> >
> > T 646.888.2354 C 516.967.0656
> >
> > ABCF-MSKCC website<
> >
>
https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility
> > >
> >
> > Adjunct Faculty
> > Center for Clinical and Translational Science
> >
> > The Rockefeller University
> > 1230 York Avenue, New York, NY 10065
> > <
>
https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g
> >
> > T 212.327.7030
> >
> > ABCF-RU website<https://www.rockefeller.edu/monoclonal/>
> >
> >
> > Please note that this e-mail and any files transmitted with it may
be
> > privileged, confidential, and protected from disclosure under
> applicable
> > law. If the reader of this message is not the intended recipient,
or
> an
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> > communication in error, please notify the sender immediately by
> replying to
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> copies and
> > backups from your computer.
> >
> >
> >
=====================================================================
> >
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I use manual columns, not FPLC.
On 8/9/21, 9:23 AM, "Antibody Discussion Forum on behalf of ABRF - Frances
Weis-Garcia" <email obscured> on behalf of <email obscured>>
wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Thanks Laura.
Trying to get away from the FPLC ...
I'll ask some "old time" purification people who may be some "old school"
methods.
On 8/9/21, 9:43 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
MabselectSure (protein A based Column). We don’t see differences and in
fact protein A works better for all Ig (in spite what literature says). The tip
was giving to me by the company that made humanization of our Star antibody:
that use mabselect for all
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of
Biomedical Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging
environment, it is crucial that our users acknowledge the following grants in
all publications (see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility,
supported in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Aug 9, 2021, at 7:50 AM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please
validate the sender's email address before clicking on links or attachments as
they may not be safe.
Yes Laura. This is exactly the stage to which I am referring.
How do you purify that 100 ml of media?
Over a Prot G resin using an FPLC?
Be well
Frances
Sent on the fly from my mobile device
On Aug 8, 2021, at 1:28 PM, David Blum <email obscured>> wrote:
Frances: I understand. This might not be the best option, but it
should
be good for processing multiple samples quickly without using the FPLC.
You could also batch bind with the resin you use now and spin down at a
speed that doesn't deform the beads. Good luck!
David
On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
Thanks David. The binding capacity is pretty low relative to standard
resins and the beads are much more expensive.
0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads
while
20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media
(formerly GE LifeSciences / Formerly Pharmacia ... boy am I old)
I know I want to use the FastFlow resin (or something similar) ...
right
now I am more looking for a process that does not involve an FPLC if I
can
help it. I have one but the person who has used it is retireing and
this
is the only thing we will use it for, so do I need it is the question.
A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each
hybridoma then line the bottles of conditioned media up on a shelf,
allowing the media to flow through a tube into a column below it and
pass
over the a prot G resin over night by gravity. Some how they set it up
so
the columns DID NOT DRY OUT. The next morning they would wash the
columns
by gravitty again in series and then elute them one at a time. Sounds
like
a great way to do a 5 to 20 at once, and I am sure I could figure this
out,
but before I spend time on this, was wondering if anyone else already
tackled this or had a better mouse trap.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
____________________________________________________________
On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum"
<
<email obscured> on behalf of <email obscured>> wrote:
Have you tried mag beads? We use them for histagged proteins at
small
scale but there are protein coupled beads too.
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqjdjH53Y$
On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
Happy Friday.
Do any of you produce and purify small amounts of the new MAbs you
make
for further characterization prior to committing to a few or one
winner?
If so, how do you do this. Currently we make 350 ml exhausted media
(hybridoma adapted to 1% FBS prescreened for ultra-low levels of
bovine IgG
+ 1 ng/ml IL6) and then purify it on a Prot G column using and
FPLC. We
get 1 – 8 mg, but I would like to scale this back and simply it if
possible
because we usually do not need that much.
Thanks.
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
<
https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqrjDVsR8$
T 646.888.2354 C 516.967.0656
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Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
<
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T 212.327.7030
ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqkt44XHc$
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Thanks Laura.
Trying to get away from the FPLC ...
I'll ask some "old time" purification people who may be some "old school"
methods.
On 8/9/21, 9:43 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
MabselectSure (protein A based Column). We don’t see differences and in
fact protein A works better for all Ig (in spite what literature says). The tip
was giving to me by the company that made humanization of our Star antibody:
that use mabselect for all
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment,
it is crucial that our users acknowledge the following grants in all
publications (see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility,
supported in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Aug 9, 2021, at 7:50 AM, Weis-Garcia, Frances/Sloan Kettering Institute
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Yes Laura. This is exactly the stage to which I am referring.
How do you purify that 100 ml of media?
Over a Prot G resin using an FPLC?
Be well
Frances
Sent on the fly from my mobile device
On Aug 8, 2021, at 1:28 PM, David Blum <email obscured>> wrote:
Frances: I understand. This might not be the best option, but it should
be good for processing multiple samples quickly without using the FPLC.
You could also batch bind with the resin you use now and spin down at a
speed that doesn't deform the beads. Good luck!
David
On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
Thanks David. The binding capacity is pretty low relative to standard
resins and the beads are much more expensive.
0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads
while
20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media
(formerly GE LifeSciences / Formerly Pharmacia ... boy am I old)
I know I want to use the FastFlow resin (or something similar) ... right
now I am more looking for a process that does not involve an FPLC if I can
help it. I have one but the person who has used it is retireing and this
is the only thing we will use it for, so do I need it is the question.
A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each
hybridoma then line the bottles of conditioned media up on a shelf,
allowing the media to flow through a tube into a column below it and pass
over the a prot G resin over night by gravity. Some how they set it up so
the columns DID NOT DRY OUT. The next morning they would wash the columns
by gravitty again in series and then elute them one at a time. Sounds like
a great way to do a 5 to 20 at once, and I am sure I could figure this out,
but before I spend time on this, was wondering if anyone else already
tackled this or had a better mouse trap.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
____________________________________________________________
On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" <
<email obscured> on behalf of <email obscured>> wrote:
Have you tried mag beads? We use them for histagged proteins at small
scale but there are protein coupled beads too.
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqjdjH53Y$
On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
Happy Friday.
Do any of you produce and purify small amounts of the new MAbs you
make
for further characterization prior to committing to a few or one
winner?
If so, how do you do this. Currently we make 350 ml exhausted media
(hybridoma adapted to 1% FBS prescreened for ultra-low levels of
bovine IgG
+ 1 ng/ml IL6) and then purify it on a Prot G column using and
FPLC. We
get 1 – 8 mg, but I would like to scale this back and simply it if
possible
because we usually do not need that much.
Thanks.
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
<
https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqrjDVsR8$
T 646.888.2354 C 516.967.0656
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The Rockefeller University
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MabselectSure (protein A based Column). We don’t see differences and in fact protein A works better for all Ig (in spite what literature says). The tip was giving to me by the company that made humanization of our Star antibody: that use mabselect for all Laura Bover, PhD (working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>) Professor Director Monoclonal Antibody Core Facility University of Texas M.D.Anderson Cancer Center Associate member of The University of Texas Graduate School of Biomedical Sciences at Houston Member of the Scientific Review Committee 2 (former Clinical Research Committee 2) Member of the Steering Committee of the Immunology Program GSBS Immunology Department/ Genomics Medicine Department 7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021 Houston- TX 77054<x-apple-data-detectors://3/0> T 713-563-3301<tel:713-563-3301> F 713-563-3276<tel:713-563-3276> <email obscured><email obscured>> In order to continue obtaining Core support in a challenging environment, it is crucial that our users acknowledge the following grants in all publications (see following text). Thank you! “We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported in part by CCSG grant number P30 CA016672.” Sent from my iPhone On Aug 9, 2021, at 7:50 AM, Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: WARNING: This email originated from outside of MD Anderson. Please validate the sender's email address before clicking on links or attachments as they may not be safe. Yes Laura. This is exactly the stage to which I am referring. How do you purify that 100 ml of media? Over a Prot G resin using an FPLC? Be well Frances Sent on the fly from my mobile device On Aug 8, 2021, at 1:28 PM, David Blum <email obscured>> wrote: Frances: I understand. This might not be the best option, but it should be good for processing multiple samples quickly without using the FPLC. You could also batch bind with the resin you use now and spin down at a speed that doesn't deform the beads. Good luck! David On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: Thanks David. The binding capacity is pretty low relative to standard resins and the beads are much more expensive. 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads while 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) I know I want to use the FastFlow resin (or something similar) ... right now I am more looking for a process that does not involve an FPLC if I can help it. I have one but the person who has used it is retireing and this is the only thing we will use it for, so do I need it is the question. A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each hybridoma then line the bottles of conditioned media up on a shelf, allowing the media to flow through a tube into a column below it and pass over the a prot G resin over night by gravity. Some how they set it up so the columns DID NOT DRY OUT. The next morning they would wash the columns by gravitty again in series and then elute them one at a time. Sounds like a great way to do a 5 to 20 at once, and I am sure I could figure this out, but before I spend time on this, was wondering if anyone else already tackled this or had a better mouse trap. Frances ZRC 1563 Office 646-888-2354 Mobile 516-967-0656
____________________________________________________________ On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" < <email obscured> on behalf of <email obscured>> wrote: Have you tried mag beads? We use them for histagged proteins at small scale but there are protein coupled beads too. https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqjdjH53Y$ On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: Happy Friday. Do any of you produce and purify small amounts of the new MAbs you make for further characterization prior to committing to a few or one winner? If so, how do you do this. Currently we make 350 ml exhausted media (hybridoma adapted to 1% FBS prescreened for ultra-low levels of bovine IgG + 1 ng/ml IL6) and then purify it on a Prot G column using and FPLC. We get 1 – 8 mg, but I would like to scale this back and simply it if possible because we usually do not need that much. Thanks. Frances Frances Weis-Garcia, Ph.D. Head Bi-Institutional Antibody and Bioresource Core Facility (RRID: CR 017691) Associate Laboratory Member Immunology Program Memorial Sloan Kettering Cancer Center 1275 York Avenue, New York, NY 10065 < https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqrjDVsR8$ T 646.888.2354 C 516.967.0656 ABCF-MSKCC website< https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqr3rckfW$ Adjunct Faculty Center for Clinical and Translational Science The Rockefeller University 1230 York Avenue, New York, NY 10065 < https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqgeETkjQ$ T 212.327.7030 ABCF-RU website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!1wQ534Un_7nW4on1VYmclsK5q_UbNqiHri00ok_zzvKRc6MMxWrL3JWPqkt44XHc$ > Please note that this e-mail and any files transmitted with it may be privileged, confidential, and protected from disclosure under applicable law. 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Yes Laura. This is exactly the stage to which I am referring. How do you purify that 100 ml of media? Over a Prot G resin using an FPLC? Be well Frances Sent on the fly from my mobile device On Aug 8, 2021, at 1:28 PM, David Blum <email obscured>> wrote: Frances: I understand. This might not be the best option, but it should be good for processing multiple samples quickly without using the FPLC. You could also batch bind with the resin you use now and spin down at a speed that doesn't deform the beads. Good luck! David On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks David. The binding capacity is pretty low relative to standard > resins and the beads are much more expensive. > >> 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads > > while > >> 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) > > I know I want to use the FastFlow resin (or something similar) ... right > now I am more looking for a process that does not involve an FPLC if I can > help it. I have one but the person who has used it is retireing and this > is the only thing we will use it for, so do I need it is the question. > > A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each > hybridoma then line the bottles of conditioned media up on a shelf, > allowing the media to flow through a tube into a column below it and pass > over the a prot G resin over night by gravity. Some how they set it up so > the columns DID NOT DRY OUT. The next morning they would wash the columns > by gravitty again in series and then elute them one at a time. Sounds like > a great way to do a 5 to 20 at once, and I am sure I could figure this out, > but before I spend time on this, was wondering if anyone else already > tackled this or had a better mouse trap. > > Frances > > ZRC 1563 > Office 646-888-2354 > Mobile 516-967-0656 > > ____________________________________________________________ > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" < > <email obscured> on behalf of <email obscured>> wrote: > > Have you tried mag beads? We use them for histagged proteins at small > scale but there are protein coupled beads too. > > https://www.thermofisher.com/order/catalog/product/88848 > > > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering > Institute <email obscured>> wrote: > >> Happy Friday. >> >> Do any of you produce and purify small amounts of the new MAbs you > make >> for further characterization prior to committing to a few or one > winner? >> If so, how do you do this. Currently we make 350 ml exhausted media >> (hybridoma adapted to 1% FBS prescreened for ultra-low levels of > bovine IgG >> + 1 ng/ml IL6) and then purify it on a Prot G column using and > FPLC. We >> get 1 – 8 mg, but I would like to scale this back and simply it if > possible >> because we usually do not need that much. >> >> Thanks. >> >> Frances >> >> >> Frances Weis-Garcia, Ph.D. >> >> Head >> Bi-Institutional Antibody and Bioresource Core Facility >> (RRID: CR 017691) >> >> Associate Laboratory Member >> Immunology Program >> >> Memorial Sloan Kettering Cancer Center >> 1275 York Avenue, New York, NY 10065 >> < > https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g >> >> T 646.888.2354 C 516.967.0656 >> >> ABCF-MSKCC website< >> > https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility >>> >> >> Adjunct Faculty >> Center for Clinical and Translational Science >> >> The Rockefeller University >> 1230 York Avenue, New York, NY 10065 >> < > https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g >> >> T 212.327.7030 >> >> ABCF-RU website<https://www.rockefeller.edu/monoclonal/> >> >> >> Please note that this e-mail and any files transmitted with it may be >> privileged, confidential, and protected from disclosure under > applicable >> law. 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Frances: I understand. This might not be the best option, but it should be good for processing multiple samples quickly without using the FPLC. You could also batch bind with the resin you use now and spin down at a speed that doesn't deform the beads. Good luck! David On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks David. The binding capacity is pretty low relative to standard > resins and the beads are much more expensive. > > > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads > > while > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) > > I know I want to use the FastFlow resin (or something similar) ... right > now I am more looking for a process that does not involve an FPLC if I can > help it. I have one but the person who has used it is retireing and this > is the only thing we will use it for, so do I need it is the question. > > A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each > hybridoma then line the bottles of conditioned media up on a shelf, > allowing the media to flow through a tube into a column below it and pass > over the a prot G resin over night by gravity. Some how they set it up so > the columns DID NOT DRY OUT. The next morning they would wash the columns > by gravitty again in series and then elute them one at a time. Sounds like > a great way to do a 5 to 20 at once, and I am sure I could figure this out, > but before I spend time on this, was wondering if anyone else already > tackled this or had a better mouse trap. > > Frances > > ZRC 1563 > Office 646-888-2354 > Mobile 516-967-0656 > > ____________________________________________________________ > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" < > <email obscured> on behalf of <email obscured>> wrote: > > Have you tried mag beads? We use them for histagged proteins at small > scale but there are protein coupled beads too. > > https://www.thermofisher.com/order/catalog/product/88848 > > > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering > Institute <email obscured>> wrote: > > > Happy Friday. > > > > Do any of you produce and purify small amounts of the new MAbs you > make > > for further characterization prior to committing to a few or one > winner? > > If so, how do you do this. Currently we make 350 ml exhausted media > > (hybridoma adapted to 1% FBS prescreened for ultra-low levels of > bovine IgG > > + 1 ng/ml IL6) and then purify it on a Prot G column using and > FPLC. We > > get 1 – 8 mg, but I would like to scale this back and simply it if > possible > > because we usually do not need that much. > > > > Thanks. > > > > Frances > > > > > > Frances Weis-Garcia, Ph.D. > > > > Head > > Bi-Institutional Antibody and Bioresource Core Facility > > (RRID: CR 017691) > > > > Associate Laboratory Member > > Immunology Program > > > > Memorial Sloan Kettering Cancer Center > > 1275 York Avenue, New York, NY 10065 > > < > https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g > > > > T 646.888.2354 C 516.967.0656 > > > > ABCF-MSKCC website< > > > https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility > > > > > > > Adjunct Faculty > > Center for Clinical and Translational Science > > > > The Rockefeller University > > 1230 York Avenue, New York, NY 10065 > > < > https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g > > > > T 212.327.7030 > > > > ABCF-RU website<https://www.rockefeller.edu/monoclonal/> > > > > > > Please note that this e-mail and any files transmitted with it may be > > privileged, confidential, and protected from disclosure under > applicable > > law. 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We start from only 100 ml, Mab select sure and get similar yield. But this is our final step for preselected candidates. We grow 2 ml supernatants of all positive clones and give those to the users for their own pilot assays (facs, blocking, WB) and then with their data and previous ELISA using 3 dilutions we decide which are the masters and purify. No more than 5. Then the user validate more. Laura Bover, PhD (working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>) Professor Director Monoclonal Antibody Core Facility University of Texas M.D.Anderson Cancer Center Associate member of The University of Texas Graduate School of Biomedical Sciences at Houston Member of the Scientific Review Committee 2 (former Clinical Research Committee 2) Member of the Steering Committee of the Immunology Program GSBS Immunology Department/ Genomics Medicine Department 7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021 Houston- TX 77054<x-apple-data-detectors://3/0> T 713-563-3301<tel:713-563-3301> F 713-563-3276<tel:713-563-3276> <email obscured><email obscured>> In order to continue obtaining Core support in a challenging environment, it is crucial that our users acknowledge the following grants in all publications (see following text). Thank you! “We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported in part by CCSG grant number P30 CA016672.” Sent from my iPhone On Aug 7, 2021, at 12:30 PM, Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: WARNING: This email originated from outside of MD Anderson. Please validate the sender's email address before clicking on links or attachments as they may not be safe. Thanks David. The binding capacity is pretty low relative to standard resins and the beads are much more expensive. 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads while 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) I know I want to use the FastFlow resin (or something similar) ... right now I am more looking for a process that does not involve an FPLC if I can help it. I have one but the person who has used it is retireing and this is the only thing we will use it for, so do I need it is the question. A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each hybridoma then line the bottles of conditioned media up on a shelf, allowing the media to flow through a tube into a column below it and pass over the a prot G resin over night by gravity. Some how they set it up so the columns DID NOT DRY OUT. The next morning they would wash the columns by gravitty again in series and then elute them one at a time. Sounds like a great way to do a 5 to 20 at once, and I am sure I could figure this out, but before I spend time on this, was wondering if anyone else already tackled this or had a better mouse trap. Frances ZRC 1563 Office 646-888-2354 Mobile 516-967-0656
____________________________________________________________ On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum" <email obscured> on behalf of <email obscured>> wrote: Have you tried mag beads? We use them for histagged proteins at small scale but there are protein coupled beads too. https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!0ywDC6YKavYActblAz3tDnLCLvIgG_uefEQ4crX1WuTTXFB3h1y313Z23adgxPJc$ On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering Institute <email obscured>> wrote: Happy Friday. Do any of you produce and purify small amounts of the new MAbs you make for further characterization prior to committing to a few or one winner? If so, how do you do this. Currently we make 350 ml exhausted media (hybridoma adapted to 1% FBS prescreened for ultra-low levels of bovine IgG + 1 ng/ml IL6) and then purify it on a Prot G column using and FPLC. We get 1 – 8 mg, but I would like to scale this back and simply it if possible because we usually do not need that much. Thanks. Frances Frances Weis-Garcia, Ph.D. Head Bi-Institutional Antibody and Bioresource Core Facility (RRID: CR 017691) Associate Laboratory Member Immunology Program Memorial Sloan Kettering Cancer Center 1275 York Avenue, New York, NY 10065 <https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!0ywDC6YKavYActblAz3tDnLCLvIgG_uefEQ4crX1WuTTXFB3h1y313Z23QcwSL5h$ > T 646.888.2354 C 516.967.0656 ABCF-MSKCC website< https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!0ywDC6YKavYActblAz3tDnLCLvIgG_uefEQ4crX1WuTTXFB3h1y313Z23Zu0LqbA$ Adjunct Faculty Center for Clinical and Translational Science The Rockefeller University 1230 York Avenue, New York, NY 10065 <https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!0ywDC6YKavYActblAz3tDnLCLvIgG_uefEQ4crX1WuTTXFB3h1y313Z23XnTDE6H$ > T 212.327.7030 ABCF-RU website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!0ywDC6YKavYActblAz3tDnLCLvIgG_uefEQ4crX1WuTTXFB3h1y313Z23cRCVF_v$ > Please note that this e-mail and any files transmitted with it may be privileged, confidential, and protected from disclosure under applicable law. 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Thanks David. The binding capacity is pretty low relative to standard resins and the beads are much more expensive. > 0.6 mg rabbit IgG / ml Pierce™ Protein G Magnetic Beads while > 20.0 mg human IgG / ml Cytiva Protein G Sepharose™ 4 Fast Flow Media (formerly GE LifeSciences / Formerly Pharmacia ... boy am I old) I know I want to use the FastFlow resin (or something similar) ... right now I am more looking for a process that does not involve an FPLC if I can help it. I have one but the person who has used it is retireing and this is the only thing we will use it for, so do I need it is the question. A lab (Ivo this was Pop Council) used to grow up a 100s of mls of each hybridoma then line the bottles of conditioned media up on a shelf, allowing the media to flow through a tube into a column below it and pass over the a prot G resin over night by gravity. Some how they set it up so the columns DID NOT DRY OUT. The next morning they would wash the columns by gravitty again in series and then elute them one at a time. Sounds like a great way to do a 5 to 20 at once, and I am sure I could figure this out, but before I spend time on this, was wondering if anyone else already tackled this or had a better mouse trap. Frances ZRC 1563 Office 646-888-2354 Mobile 516-967-0656
____________________________________________________________
On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
Have you tried mag beads? We use them for histagged proteins at small
scale but there are protein coupled beads too.
https://www.thermofisher.com/order/catalog/product/88848
On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Happy Friday.
>
> Do any of you produce and purify small amounts of the new MAbs you make
> for further characterization prior to committing to a few or one winner?
> If so, how do you do this. Currently we make 350 ml exhausted media
> (hybridoma adapted to 1% FBS prescreened for ultra-low levels of bovine
IgG
> + 1 ng/ml IL6) and then purify it on a Prot G column using and FPLC. We
> get 1 – 8 mg, but I would like to scale this back and simply it if
possible
> because we usually do not need that much.
>
> Thanks.
>
> Frances
>
>
> Frances Weis-Garcia, Ph.D.
>
> Head
> Bi-Institutional Antibody and Bioresource Core Facility
> (RRID: CR 017691)
>
> Associate Laboratory Member
> Immunology Program
>
> Memorial Sloan Kettering Cancer Center
> 1275 York Avenue, New York, NY 10065
>
<https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g>
> T 646.888.2354 C 516.967.0656
>
> ABCF-MSKCC website<
>
https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility
> >
>
> Adjunct Faculty
> Center for Clinical and Translational Science
>
> The Rockefeller University
> 1230 York Avenue, New York, NY 10065
>
<https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g>
> T 212.327.7030
>
> ABCF-RU website<https://www.rockefeller.edu/monoclonal/>
>
>
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Have you tried mag beads? We use them for histagged proteins at small scale but there are protein coupled beads too. https://www.thermofisher.com/order/catalog/product/88848 On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Happy Friday. > > Do any of you produce and purify small amounts of the new MAbs you make > for further characterization prior to committing to a few or one winner? > If so, how do you do this. Currently we make 350 ml exhausted media > (hybridoma adapted to 1% FBS prescreened for ultra-low levels of bovine IgG > + 1 ng/ml IL6) and then purify it on a Prot G column using and FPLC. We > get 1 – 8 mg, but I would like to scale this back and simply it if possible > because we usually do not need that much. > > Thanks. > > Frances > > > Frances Weis-Garcia, Ph.D. > > Head > Bi-Institutional Antibody and Bioresource Core Facility > (RRID: CR 017691) > > Associate Laboratory Member > Immunology Program > > Memorial Sloan Kettering Cancer Center > 1275 York Avenue, New York, NY 10065 > <https://www.google.com/maps/search/1275+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g> > T 646.888.2354 C 516.967.0656 > > ABCF-MSKCC website< > https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility > > > > Adjunct Faculty > Center for Clinical and Translational Science > > The Rockefeller University > 1230 York Avenue, New York, NY 10065 > <https://www.google.com/maps/search/1230+York+Avenue,+New+York,+NY+10065?entry=gmail&source=g> > T 212.327.7030 > > ABCF-RU website<https://www.rockefeller.edu/monoclonal/> > > > Please note that this e-mail and any files transmitted with it may be > privileged, confidential, and protected from disclosure under applicable > law. If the reader of this message is not the intended recipient, or an > employee or agent responsible for delivering this message to the intended > recipient, you are hereby notified that any reading, dissemination, > distribution, copying, or other use of this communication or any of its > attachments is strictly prohibited. If you have received this > communication in error, please notify the sender immediately by replying to > this message and deleting this message, any attachments, and all copies and > backups from your computer. > > > ===================================================================== > > Please note that this e-mail and any files transmitted from > Memorial Sloan Kettering Cancer Center may be privileged, > confidential, > and protected from disclosure under applicable law. If the reader of > this message is not the intended recipient, or an employee or agent > responsible for delivering this message to the intended recipient, > you are hereby notified that any reading, dissemination, distribution, > copying, or other use of this communication or any of its attachments > is strictly prohibited. If you have received this communication in > error, please notify the sender immediately by replying to this > message > and deleting this message, any attachments, and all copies and backups > from your computer. > > ―― > View topic http://list.abrf.org/r/topic/6Md8fJOjSbIvYb2uRBONUs > Leave group <email obscured>?Subject=Unsubscribe > >
Happy Friday. Do any of you produce and purify small amounts of the new MAbs you make for further characterization prior to committing to a few or one winner? If so, how do you do this. Currently we make 350 ml exhausted media (hybridoma adapted to 1% FBS prescreened for ultra-low levels of bovine IgG + 1 ng/ml IL6) and then purify it on a Prot G column using and FPLC. We get 1 – 8 mg, but I would like to scale this back and simply it if possible because we usually do not need that much. Thanks. Frances Frances Weis-Garcia, Ph.D. Head Bi-Institutional Antibody and Bioresource Core Facility (RRID: CR 017691) Associate Laboratory Member Immunology Program Memorial Sloan Kettering Cancer Center 1275 York Avenue, New York, NY 10065 T 646.888.2354 C 516.967.0656 ABCF-MSKCC website<https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility> Adjunct Faculty Center for Clinical and Translational Science The Rockefeller University 1230 York Avenue, New York, NY 10065 T 212.327.7030 ABCF-RU website<https://www.rockefeller.edu/monoclonal/> Please note that this e-mail and any files transmitted with it may be privileged, confidential, and protected from disclosure under applicable law. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any reading, dissemination, distribution, copying, or other use of this communication or any of its attachments is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting this message, any attachments, and all copies and backups from your computer.
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Hi Frances, I'm not sure what stage you're at with screening these hybridomas, but a possibility could be that you're picking up on "contaminating" antibody from antibody diffused from a different hybridoma in the methylcellulose in the proximity of the chosen clone when the clones are picked. If this is the first screen of these hybridomas after clone picking that might be the case. We've found that with several passages of the clones the non-specific antibody signal goes away. Are the IgGs and IgMs specific to the target with an isotype specific secondary in an indirect ELISA? Just a thought. Thanks, Kurt Kurt Christensen Lab Director Protein and Monoclonal Antibody Production Core Web Page: https://www.bcm.edu/centers/cancer-center/research/shared-resources/antibody-based-proteomics/recombinant-protein-monoclonal-antibody iLab Requests: https://bcm.corefacilities.org/account/login Office: 713-798-2325 Lab:713-798-2327 Email: <email obscured> Baylor College of Medicine Margaret M. Alkek Building for Biomedical Research, Room R551E, Mail Stop BCM929 1 Baylor Plaza Houston, Texas 77030
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Weis-Garcia, Frances/Sloan Kettering Institute
Sent: Tuesday, July 27, 2021 10:32 PM
To: ABRF Antibody Discussion Group <email obscured>>
Subject: [ABRF Ab Forum] Hybridoma sups scoring positive for IgG and IgM
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://urldefense.proofpoint.com/v2/url?u=https-3A__www.mskcc.org_research-2Dadvantage_core-2Dfacilities_monoclonal-2Dantibody-2Dcore-2Dfacility&d=DwIFaQ&c=ZQs-KZ8oxEw0p81sqgiaRA&r=_dqqB8TU9vN7Hm15OCeZVQ&m=Cq-Nv9S-ZYzO9JkVdbbaJW231UrAb7laQwd2Y5diEXQ&s=-g9lJvrqUJEPum7Vtra94yqlTuEf-HIOAf7jJCf5WGg&e=
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://urldefense.proofpoint.com/v2/url?u=https-3A__www.rockefeller.edu_monoclonal_&d=DwIFaQ&c=ZQs-KZ8oxEw0p81sqgiaRA&r=_dqqB8TU9vN7Hm15OCeZVQ&m=Cq-Nv9S-ZYzO9JkVdbbaJW231UrAb7laQwd2Y5diEXQ&s=L04n0wmp7XTkxI7aCThBxnecYhaiODn1-OBRmgje-OQ&e=
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I agree, if we were purifying.
All we want to see is if they are both there with a different technique ... and
I want to SEE the HC band ... not just color in a well __
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/29/21, 9:51 AM, "Antibody Discussion Forum on behalf of Carnahan, Robert"
<email obscured> on behalf of <email obscured>> wrote:
I guess my only two cents is we had way better luck with capture select
resins than Protein L. Can get kappa or lambda specific resins.
Good luck.
Rob
From: Antibody Discussion Forum <email obscured>> on behalf of
Bover,Laura <email obscured>>
Date: Thursday, July 29, 2021 at 8:43 AM
To: <email obscured> <email obscured>>
Subject: Re: [ABRF Ab Forum] Re: [EXT] Hybridoma sups scoring positive for
IgG and IgM
I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
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Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to
get to confirm the ELISA results. Next set would be to subclone a few to make
sure they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of
Biomedical Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
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<email obscured><email obscured>>
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Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
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Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we
have secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG
and 1 IgM. We have not subcloned yet … but these cultures did come from a
ClonePix run.
Thoughts ???
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__https%3A%2F%2Fwww.mskcc.org%2Fresearch-advantage%2Fcore-facilities%2Fmonoclonal-antibody-core-facility__%3B!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai%24&data=04%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cf0b6fb49e42b4e92aaf708d95296e5ff%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C1%7C637631630326968143%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=BxaTRTc2GKSCdyN%2Be4%2FLdzNhZY6Bv3iYM2YHDkTA6ho%3D&reserved=0
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Adjunct Faculty
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T 212.327.7030
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I guess my only two cents is we had way better luck with capture select resins
than Protein L. Can get kappa or lambda specific resins.
Good luck.
Rob
From: Antibody Discussion Forum <email obscured>> on behalf of
Bover,Laura <email obscured>>
Date: Thursday, July 29, 2021 at 8:43 AM
To: <email obscured> <email obscured>>
Subject: Re: [ABRF Ab Forum] Re: [EXT] Hybridoma sups scoring positive for IgG
and IgM
I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering Institute
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
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Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment, it
is crucial that our users acknowledge the following grants in all publications
(see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported
in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering Institute
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
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Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__https%3A%2F%2Fwww.mskcc.org%2Fresearch-advantage%2Fcore-facilities%2Fmonoclonal-antibody-core-facility__%3B!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai%24&data=04%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cf0b6fb49e42b4e92aaf708d95296e5ff%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C1%7C637631630326968143%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=BxaTRTc2GKSCdyN%2Be4%2FLdzNhZY6Bv3iYM2YHDkTA6ho%3D&reserved=0
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Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
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I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering Institute
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment, it
is crucial that our users acknowledge the following grants in all publications
(see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported
in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering Institute
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai$
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaFYFUkAw$
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Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment,
it is crucial that our users acknowledge the following grants in all
publications (see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility,
supported in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please validate
the sender's email address before clicking on links or attachments as they may
not be safe.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
Frances
Frances Weis-Garcia, Ph.D.
Head
Bi-Institutional Antibody and Bioresource Core Facility
(RRID: CR 017691)
Associate Laboratory Member
Immunology Program
Memorial Sloan Kettering Cancer Center
1275 York Avenue, New York, NY 10065
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai$
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
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