Novel homogeneous enzyme immunoassay: chitinase activity enhancement immunoassay (CAEIA).
A novel homogeneous enzyme immunoassay has been developed that takes advantage of the observation that the activity of a bacterial chitinase could be enhanced by the binding of a ligand conjugate of a monoclonal antibody to the enzyme (L-MAbchi). The enhancement was suppressed by an antiligand antibody (AbL). Free ligand molecules competed for AbL combining with L-MAbchi, and the suppression of the enhancement was reduced. Thus the concentration of free ligand could be estimated. Biotin was used as a ligand in this model experiment. When this monoclonal antibody was conjugated with a ligand (e.g., biotin), we furthermore found that the enhancement could be suppressed by an antiligand antibody (AbL). When free ligand molecules are added to the system, competition with L-MAbchi results to reduce the suppression of enhancement.
Sekine
S
S
Department of Biosystem Science, Graduate School of Science and Technology, Niigata University, Japan.
Watanabe
T
T
Tanaka
H
H
D000118:Acetylglucosaminidase / Q000097:blood; D000911:Antibodies, Monoclonal; D001407:Bacillus / Q000201:enzymology; D001667:Binding, Competitive / Q000276:immunology; D001710:Biotin / Q000032:analysis / Q000276:immunology; D002688:Chitinases / Q000097:blood / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques
D000911:Antibodies, Monoclonal; D001710:Biotin; D002688:Chitinases; D000118:Acetylglucosaminidase