Production of Potent Anti-Australia Antigen Sera of High Specificity and Sensitivity in Goats

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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 p H treatment, isopycnic centrifugation two times in CsCl, and rate zonal centrifugation in sucrose; (ii) same as procedure i, with the exception of the low p H 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.