Detection of IgG in supernatants of pokeweed mitogen-stimulated human lymphocyte cultures by one step solid-phase radioimmunoassay (SPRIA).
A one step solid phase radioimmunoassay is used as a simple and reproducible method of detection and quantitation of IgG produced by human PBL after stimulation with PWM. Modifications of culture conditions are necessary to make culture supernatants suitable for this assay. Pulsing with PWM must be performed in serum-supplemented culture medium for 4-5 days. After thorough washing, medium is then replaced with serum-free medium. Under these conditions, synthesis and secretion of IgG continues for at least 9 days. The amount of IgG produced by 10(6) normal adult PBL as detected in this system is 0.77 +/- 0.47 micrograms. No close correlation between cell proliferation and IgG synthesis was observed.
D003470:Culture Media; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis*; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology; D011043:Pokeweed Mitogens / Q000494:pharmacology; D011863:Radioimmunoassay / Q000379:methods*
D003470:Culture Media; D007074:Immunoglobulin G; D011043:Pokeweed Mitogens