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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