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The first is Gray et al.1995 J.Immuno.Meth.182:155, Secretion capture and
report web: use of affinity derivatized agarose microdroplets for the selection of
hybridoma cells. The second paper is Kenney et al. and should be out in
Bio/Technology in August.
These papers describe work some of you have seen in posters at ISAC. We had to
stretch a little to get a SCRWy acronym, but the assay is pretty cool. It's some
developments on a technique described by Jim Weaver at MIT and commercialized by
One Cell Systems in Cambridge MA wherein secreted products of individual cells are
specifically captured in a ~30um droplet which surrounds each cell and reported
with fluorescent antibodies or antigens or whatever ligand you build. The
preparation can be pretty easily flow analyzed, sorted or cloned. You don't seem
to hurt the cells, the assay shows very good resolution of secreted antibody
specificity, reasonable quantitation of hypersecretory phenotypes, and pretty good
"rare event" (~1%) sensitivity.
The bottom line for us is that this is the way we make antibodies from now on.
We do a normal immunization and fusion, then we screen and clone in a day by
SCRWing a bulk culture of HAT-selected cells. I have some experience with doing
cell surface staining for hybridoma screening and agree that it can be made to
work, but I've found it to be inconsistent at best. We have always light-scatter
flow-cloned our hybridomas using both Coulter's or BD's cloning devices. They work
great and I am amazed at how under-utilized they are in most facilities.
Limit-dilution lost it's charm for me a long time back. The SCRW technique is a
very powerful addition (please excuse the crowing) in that it quantitatively and
qualitatively measures secretion directly.
I might state the obvious-it has lots of potential for other secretion assays
beyond hybridomas.
My apologies for the belated response. My lab was recently devoured like a
ripe watermelon during the Roche acquisition of Syntex, and I have enjoyed the fate
of the little black pit that I am. I am no longer available at
Jack.Dunne@Syntex.com, and so have been reading these postings over my colleague
Laura.Chiu@Syntex.com's shoulder somewhat sporadically. Old and new friends are
invited to contact me at home:
Uncle Jack Dunne
1210 Payne Dr
Los Altos, CA 94024
415 988 0628
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