Passive Immune Hemolysis: Titration of Hemolytic Anti-Protein Antibody Using Bis-Diazotized-Benzidine to Couple Antigen to Erythrocytes.

Publisher Website
Google Scholar
Abstract
Summary By modification of reagents and conduct of reaction, the well-known technique for coupling protein antigens to erythrocytes with bis-diazotized-benzidine, already useful in detecting specific antibody by hemagglutination, can be adapted to quantitative titration of hemolytic antibody by passive immune hemolysis. This paper describes the details of preparation of the indicator cells and shows the specificity of their reaction. When antisera against a protein antigen are prepared in guinea pigs, hemolysis in this system will occur only in the presence of the complement-fixing, cytotoxic, hemolytic 7Sγ2 antibody. The reproducibility of the method described was determined by 23 consecutive titrations of a standard hemolytic antiserum: the mean value was 1124 50% hemolysin units with a standard deviation of ± 166 (= 14.8%). The sensitivity of the present technique was found to be 1/10th to l/20th as sensitive as hemagglutination upon comparative analysis of isolated γ2 antibody. This is an improvement over the sensitivity of current immunoelectrophoretic assays for the γ2 antibody.

This publication has 0 references indexed in Scilit: