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
> > > > <
> > >
> >
>
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> > > >
> > > > 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
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> > > >
> > > > The Rockefeller University
> > > > 1230 York Avenue, New York, NY 10065
> > > > <
> > >
> >
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> > > >
> > > > T 212.327.7030
> > > >
> > > > ABCF-RU website<
>
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