Monoclonal antibodies against soybean Bowman-Birk inhibitor recognize the protease-reactive loops.
Monoclonal antibodies against soybean Bowman-Birk protease inhibitor (BBI) have been generated and used to detect and quantify BBI in foods, soybean germplasm, and animal tissues and fluids. The purpose of this study was to determine the recognition sites of two monoclonal antibodies to BBI (mAb 238 and mAb 217) in relation to the protease-inhibitory sites of BBI. The results showed that (1) the binding of mAb 238 can be blocked by trypsin and that of mAb 217 by chymotrypsin; (2) the trypsin or chymotrypsin inhibitory activities of BBI are blocked by mAb 238 or mAb 217, respectively; and (3) mAb 238 failed to recognize a tryptic loop mutant BBI variant and mAb 217 was unable to bind a chymotryptic loop mutant BBI variant. These findings demonstrate that the epitopes recognized by mAb 238 and mAb 217 reside, at least in part, in the tryptic and chymotryptic loops of BBI, respectively.
Mao
Yifan
Y
Genencor International, Inc., 925 Page Mill Road, Palo Alto, CA 94304, USA.
Lai
Cindy
C
Vogtentanz
Gudrun
G
Schmidt
Brian
B
Day
Tony
T
Miller
Jeff
J
Brandon
David L
DL
Chen
Dan
D
D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D001666:Binding Sites, Antibody; D002918:Chymotrypsin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D014357:Trypsin / Q000737:chemistry* / Q000494:pharmacology*; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000737:chemistry* / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D002918:Chymotrypsin; D014357:Trypsin