Use of Recycled Antibodies for the Immunometric Assay of Hepatitis B Surface Antigen

The mechanism of the hook effect in a two site immunometric assay of hepatitis B surface antigen (HBsAg) was studied using a novel method. The results showed that HBsAg dissociated from immobilized anti-HBsAg at low as well as high doses of antigen (Ag). This probably was not due to lack of immobilized antibody (Ab) or dissociation of HBsAg non-specifically bound to the immobilized Ab. The dissociated Ag sequestered the labelled Ab in the second reaction and contributed to the decrease in percent binding (B%) of the assay and thus the hook effect. The dissociated Ag could contaminate the recovered pooled labelled Ab in recycled assay and give rise to high non-specific binding. Prolonged washing decreased the side reaction but also the B% of the assay.

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Use of Recycled Antibodies for the Immunometric Assay of Hepatitis B Surface Antigen

Cited by 2 publications

References 2 publications

Effect of serum, solvent and detergent on antibody immobilized on polymeric substrates and its influence on antigen-antibody interaction

Effect of serum, solvent and detergent on antibody immobilized on polymeric substrates and its influence on antigen-antibody interaction

Dissociation of Hepatitis B Surface Antigen (HBsAg) from Immobilised Anti—HBsAg Antibody in a Two Site Immunoradiometric Assay: Its Relevance to Hook Effect and Recycled Assay

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