Purification of soluble complement-fixing antigens from two Burkitt's lymphoma cell lines. Conditions affecting the stability and recovery of antigens and antibodies.
Soluble Epstein--Barr virus (EBV) associated antigens of P3HR-1 and Raji cell lines, which fix complement in the presence of human sera containing antibody against EBV--VCA antigens, were partially purified by fractionating centrifuged cell lysates by the method of immunoadsorbents or by precipitation with ammonium sulphate or glycine--HCl. Purification conditions were limited by the instability of both antigen and antibody activity. The antibody was inactivated under all the conditions commonly used to dissociate antigen--antibody complexes. The antigen was inactivated at pH 2-3, and was irreversibly bound to Sephadex G-200 conjugates which had not been exposed to large excesses of serum.
D000327:Adsorption; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000302:isolation & purification*; D002460:Cell Line; D011232:Chemical Precipitation; D003168:Complement Fixation Tests; D004591:Electrophoresis, Polyacrylamide Gel; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D006863:Hydrogen-Ion Concentration
D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral