Screening anti-human IgG for colloidal gold conjugate with dot immunogold filtration assay / 中国免疫学杂志

ABSTRACT

Objective:To screen the appropriate anti-human IgG (secondary antibody) for conjugating with colloidal gold by dot immunogold filtration assay,and put forward the method of using multiple evaluation indicators for secondary antibody comparing and analyzing. Methods:Three different secondary antibodies A,B and C from three different companies were screened. Firstly the titration was used to determine the optimal reaction conditions. Then three secondary antibodies were conjugated to the colloidal gold,tested with a dot immunogold filtration assay for hydatidosis specific antibody detection. The optimal conjugated secondary antibody were compared with the standard indirect enzyme-linked immunosorbent assay. Results:The optimal pH of three secondary antibodies were 8. 5,the binding capacity were 38. 4,24 and 19. 2μg /ml colloidal gold. According to the comprehensive evaluation,the diagnostic effects of sec-ondary antibody B was better than others. Its diagnostic effects in dot immunogold filtration assay was compared with enzyme-linked im-munosorbent assay. The Kappa was 0. 895(P<0. 05) and showed the two methods were in good agreement. Conclusion:The appropriate secondary antibody for conjugating with colloidal gold could be screened by dot immunogold filtration assay and the evaluation indicators which put forward by this study,the screening method would have an important reference value for all kinds of colloidal gold test kit to screen the suitable secondary antibody.