Comparison Method of Reactivity of Anti-Human Immunoglobulin Reagents for Flow Cytometry

Won, Dong Il

Abstract

BACKGROUND
In flow cytometry, the verification of the anti-human immunoglobulin (anti-Ig) reactivity is important. The author has devised a simple protocol for this. METHODS: As a staining verification control, red blood cells (RBCs) were used. IgG- or IgM- coated RBCs were made using group O or group B serum, respectively. Ani-Ig reactivity in each serially diluted serum for immunoglobulin coating and correlation between the amount of reacting anti-Ig on RBCs and measured values were investigated. In one case of parallel tests for two anti-Ig reagents, author's method was compared with the B cell gating method. RESULTS: Both the test/control mean channel fluorescence ratio and percent fluorescence had significant linear correlations with the amount of the reacting anti-IgG on RBCs. As well as B cell gating, the author's method seemed to also clearly discriminate the reactivity of two anti-IgG reagents. CONCLUSIONS: Using immunoglobulin-coated RBCs as a staining verification control, the parallel test for two anti-IgG reagents could be performed more conveniently and objectively than the B cell gating method.