Purification of acute phase anti-hepatitis A virus (HAV) IgM and development of an IgM solid-phase radioimmunoassay for the detection of HAV.
A simple two-step procedure for the purification of IgM from acute phase hepatitis A infected chimpanzee serum has been developed. The procedure involves the precipitation of the euglobulin fraction with boric acid and the fractionation of the resuspended material on Sephacryl S-300 (Pharmacia Fine Chemicals, Piscataway, NJ). The purified IgM was used to develop a solid-phase radioimmunoassay (SPRIA) in which IgM was used as capture antibody and iodinated IgM was used as a probe. This assay was compared with a conventional IgG-based SPRIA. The IgM-based assay resulted in a superior response (P/N) for the detection of antigen in crude preparations; however, endpoint analyses demonstrated similar sensitivities.
D000208:Acute Disease; D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification*; D002143:Callitrichinae; D002850:Chromatography, Gel; D006506:Hepatitis A / Q000175:diagnosis* / Q000276:immunology; D006507:Hepatovirus / Q000032:analysis / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification* / Q000378:metabolism; D008099:Liver / Q000032:analysis; D002679:Pan troglodytes; D011863:Radioimmunoassay / Q000379:methods
D000914:Antibodies, Viral; D007075:Immunoglobulin M