Production and characterization of monoclonal antibodies to rat liver arginase.
Rat liver arginase was purified and five monoclonal antibodies were produced by fusion of spleen cells from a Balb/c mouse and the myeloma cell line P3-X36-Ag-U1. One, R2D19, of five antibodies belonged to the IgG2a subclass, the other four, R1D81, R1G11, R2E10, and R2G51, were of the IgG1 type. The R1D81 cross-reacted with human liver arginase. This antibody inhibited the arginase activity, competing with arginine. These results suggest that R1D81 binds to the catalytic site of arginase. The R2D19 also inhibited the enzyme activity but acted as a noncompetitive inhibitor. With the use of R1D81 and a polyclonal anti-human liver arginase antibody conjugated with alkaline phosphatase, a sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the quantification of human arginase. Specificity of monoclonal antibodies for rat liver arginase was examined by means of the sandwich ELISA. Eight pairs of monoclonal antibodies could form a sandwich with the arginase. Only the R2E10 could be used for both the first and the second antibody in the sandwich system. In other cases, monoclonal antibodies could not be interchanged between solid and liquid phase.
Isoai
A
A
Research Center, Asahi Glass Co., Ltd., Kanagawa.
Harie
S
S
Uchida
K
K
Hirai
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D001119:Arginase / Q000276:immunology*; D001120:Arginine / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006041:Goats / Q000276:immunology; D006801:Humans; D007136:Immunoglobulins; D008099:Liver / Q000201:enzymology*; D051381:Rats; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D001120:Arginine; D001119:Arginase