We've had some success using immobilized antigens to pan out B cells
reactive to them. It's a bit trickey and requires you get the B cells
while they are in the periphery and still have sufficient amounts of
membrane Ig on them. Mostly the numbers of B cells in the periphery peak
7-10 days after immunization and are pretty much gone by 2 weeks. If
you're not specifically immunizing people then it could be very tough to
get the B cells that you need.
Immobilizing the antigen on a sterile petri plate works. Also, if its
possible to immobilize your antigen on a tissue culture plate you could
try doing this directly onto the wells that you will then grow the cells
in.
As for obtainijng the B cells in a fairly purified form, the usual methods
(ficoll or percoll gradients) should suffice.
Jerry Pier
In article <43iqmb$he1 at moe.cc.emory.edu>, mediml at pop3.service.emory.edu wrote:
> Any suggestions or references on how to isolate human lymphocytes from
> peripheral blood that express immunoglobulins reactive against a
> defined antigen? If such lymphocytes are not part of a neoplastic
> response, is their identification and isolation a figment of my
> immagination?
>> Thanks,
>> Ira
