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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