The Protein Journal

, Volume 24, Issue 5, pp 275–282 | Cite as

Monoclonal Antibodies Against Soybean Bowman-Birk Inhibitor Recognize the Protease-Reactive Loops

  • Yifan Mao
  • Cindy Lai
  • Gudrun Vogtentanz
  • Brian Schmidt
  • Tony Day
  • Jeff Miller
  • David L. Brandon
  • Dan Chen
Article

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.

Keywords

Bowman-Birk protease inhibitor chymotrypsin epitope monoclonal antibody trypsin 

Abbreviations

BBI

Bowman-Birk protease inhibitor

mAb

monoclonal antibody

SBTI

soybean trypsin inhibitor

AAPF

succinyl-ala-ala-pro-phe-paranitroanilide

AAPR

succinyl-ala-ala-pro-arg-paranitroanilide

ELISA

enzyme-linked immunosorbent assay

PBS

phosphate-buffered saline

SDS-PAGE

sodium dodecylsulfate polyacrylamide gel electrophoresis.

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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Yifan Mao
    • 1
  • Cindy Lai
    • 1
  • Gudrun Vogtentanz
    • 1
  • Brian Schmidt
    • 1
  • Tony Day
    • 1
  • Jeff Miller
    • 1
  • David L. Brandon
    • 2
    • 3
  • Dan Chen
    • 1
    • 3
  1. 1.Genencor International, Inc.Palo AltoUSA
  2. 2.Western Regional Research CenterUS Department of Agriculture, Agricultural Research ServiceAlbanyUSA
  3. 3.Dan Chen M.D.Los Altos

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