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A big advantage of this method is that you can start antibody production
immediately. If you have pristane-primed mice available, you can
sort in a single day (NON-STERILE if you wish!) and inject the sorted
cells directly into mice. This doesn't guarantee clonal stability
(since you haven't cloned) but gets you lots of antibody in the least
time (saving the weeks needed for limiting dilution enrichment, screening,
limiting dilution cloning, re-screening, and clone expansion).
I'd be interested in the basis for Rob Chervenak's statement:
"in our experience, there is little correlation between cell surface Ig and
the level of secretion of Ab by hybridoma cells". Was I just lucky?
In message Fri, 30 Jun 1995 17:22:06 -0400 (edt),
nbenson@nervm.nerdc.ufl.edu (Neal Benson) writes:
> One of my lab's users has several hybridoma cell lines that they would
> like to re-clone to improve antibody production. It was suggested that
> they use cell sorting to select the "high-expressers". They propose to
> label the cells with a FITC-labeled anti-mouse IgG (subclass specific),
> then sort those in the upper tail of the resulting fluorescence histogram.
>
> In my limited knowledge of this area, I seem to have the impression that
> expression of antibody on the cell surface does not necessarily correspond
> to the quantity of antibody secreted from a hybridoma cell. I'd be very
> interested to obtain opinions as to whether sorting of these cells based
> on high surface expression is a reasonable thing to do for improvement of
> antibody production.
>
> Thanks in advance,
> Neal
> =======================================================================
> Neal A. Benson Tel: (904) 392-0008
> University of Florida Fax: (904) 392-4693
> Department of Pathology Email: nbenson@nervm.nerdc.ufl.edu
> Box 100275
> Gainesville, FL 32610-0275 USA
>
/*- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Eric Martz, Professor of Immunology emartz@microbio.umass.edu
Dept Microbiology Voice: 413-545-2325 FAX: 413-545-1578
Morrill IVN 203, Box 35720, Univ Massachusetts, Amherst MA 01003-5720
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