title: Production of potent anti-Australia antigen sera of high specificity and sensitivity in goats.
abstract: Potent antisera of high specificity and sensitivity were produced, in goats, to purified Australia antigen (Au). The antigen was prepared by one of three methods: (i) pelleting, low pH treatment, isopycnic centrifugation two times in CsCl, and rate zonal centrifugation in sucrose; (ii) same as procedure i, with the exception of the low pH treatment; or (iii) twice banding in CsCl by using a BXIV batch-type zonal centrifuge rotor with subsequent preparative Pevikon electrophoresis. The goat anti-Au sera contained high levels of precipitating antibody as tested by immunodiffusion in agar gel and discontinuous counterimmunoelectrophoresis (DCIE) as well as specific complement-fixing antibody and could be used for routine screening of sera for Au without prior adsorption with Au-negative normal human serum (NHS). Identification of 66 of 70 positive specimens (94.3%) in a panel of 98 coded sera (49 duplicates) with 100% reproducibility was made by using one of the goat anti-Au sera at a dilution of 1:16 in the DCIE method. No false positives were recorded. Low levels of antibody against NHS components were effectively removed by a single adsorption with glutaraldehyde cross-linked NHS.
mesh_terms: D000327:Adsorption; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D000917:Antibody Formation; D000918:Antibody Specificity; D002499:Centrifugation, Density Gradient; D006041:Goats / Q000276:immunology*; D006168:Guinea Pigs / Q000276:immunology; D006511:Hepatitis B Antigens / Q000302:isolation & purification; D007106:Immune Sera; D007122:Immunoelectrophoresis; D007456:Iodine Isotopes; D008854:Microscopy, Electron; D011863:Radioimmunoassay
chemical_list: D000914:Antibodies, Viral; D006511:Hepatitis B Antigens; D007106:Immune Sera; D007456:Iodine Isotopes