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<
> > > >
> > >
> >
>
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> > > > >
> > > >
> > > > Adjunct Faculty
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> > > >
> > > > The Rockefeller University
> > > > 1230 York Avenue, New York, NY 10065
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> > >
> >
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>
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