Probing the Soybean Bowman–Birk Inhibitor Using Recombinant Antibody Fragments
| Autor: | Julien Muzard, Conor Fields, James J. O’Mahony, Gil U. Lee |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Trypsin inhibitor
Molecular Sequence Data Epitope law.invention Magnetics Mice 03 medical and health sciences law medicine Animals Chymotrypsin Trypsin Amino Acid Sequence Cloning Molecular Immunoglobulin Fragments Soy protein Trypsin Inhibitor Bowman-Birk Soybean 030304 developmental biology Immunoassay 0303 health sciences Hybridomas biology medicine.diagnostic_test Chemistry 030302 biochemistry & molecular biology General Chemistry Molecular biology Recombinant Proteins 3. Good health biology.protein Recombinant DNA Binding Sites Antibody Soybeans Soybean Proteins General Agricultural and Biological Sciences Single-Chain Antibodies medicine.drug |
| Zdroj: | Journal of Agricultural and Food Chemistry |
| ISSN: | 1520-5118 0021-8561 |
| Popis: | The nutritional and health benefits of soy protein have been extensively studied over recent decades. The Bowman-Birk inhibitor (BBI), derived from soybeans, is a double-headed inhibitor of chymotrypsin and trypsin with anticarcinogenic and anti-inflammatory properties, which have been demonstrated in vitro and in vivo. However, the lack of analytical and purification methodologies complicates its potential for further functional and clinical investigations. This paper reports the construction of anti-BBI antibody fragments based on the principle of protein design. Recombinant antibody (scFv and diabody) molecules targeting soybean BBI were produced and characterized in vitro (K(D)~1.10(-9) M), and the antibody-binding site (epitope) was identified as part of the trypsin-specific reactive loop. Finally, an extremely fast purification strategy for BBI from soybean extracts, based on superparamagnetic particles coated with antibody fragments, was developed. To the best of the authors' knowledge, this is the first report on the design and characterization of recombinant anti-BBI antibodies and their potential application in soybean processing. |
| Databáze: | OpenAIRE |
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