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/>
> > >
> > >
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