A new approach for producing polyclonal antibodies using impure antigens.
A new convenient approach has been designed to produce polyclonal antibodies (PcAb). The approach is based on the principle of the immunoglobulin (Ig) class switch in the immune response. We produced six different antibodies (Ab) against calcineurin A subunit (CNA). CNA, His-tagged calcineurin A subunit (His-CNA), single chain calcineurin (CNB-CNA) and single chain calcineurin-calmodulin complex (CaM-CNB-CNA) were expressed in Escherichia coli (E. coli) BL21 strain, and they were used to immunize male BALB/c mice. These Ab were examined by enzyme-linked immunosorbent assay (ELISA) and western blot analysis. The results clearly demonstrated that the specificity of the Ab produced by different protein samples was much higher than that of the Ab produced by a single sample. We used CNA, CaM-CNB-CNA and His-CNA to immunize mice in turn and obtained monospecific PcAb against CNA fortunately by our new approach. Remarkably, our approach not only offered a simple and general alternative to other methods for producing PcAb described previously, but also disclosed a novel process of immunization that could be used to produce monoclonal antibodies (mAb).
Yang
Chenggang
C
Department of Biochemistry and Molecular Biology, Beijing Normal University, Beijing Key Laboratory, Beijing 100875, China.
Song
Hongtao
H
Wang
Yanli
Y
Peng
Fei
F
Wei
Qun
Q
D000818:Animals; D000906:Antibodies / Q000008:administration & dosage*; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology*; D019703:Calcineurin / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D007114:Immunization / Q000379:methods; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D021122:Protein Subunits / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000906:Antibodies; D000941:Antigens; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D019703:Calcineurin