Immunoprecipitation of pathogen proteins with Human serum? - (Mar/22/2006 )

Hi,

I would like to identify all the bacterial proteins from a pathogen that are generating an antibody response in humans. I have serum from patients available and was wondering if there is any reason why I can t use the serum to immunoprecipitate proteins from a total cell lysate of the pathogen???

I think the pathogen specific antibodies will be quite low in the serum as the patients are in remmission ie dont have active disease. I was therefore thinking of purifying Ig's from the serum beforehand, is this a good idea.

Thanks

Mark

total cell lysate? then you are only looking for proteins that are not secreted...will you use a native lysis protocol, with protease inhibitors and such?

if you are looking for antigens only, then I would not use whole serum...your task in weeding through the pile of proteins you get will be hard enough so the idea of using IgG, I think, is good