Probing the soybean Bowman-Birk inhibitor using recombinant antibody fragments.
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.
Bionanosciences, UCD Centre for Nanomedicine, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland. julien.muzard@ucd.ie
Julien
J
Muzard
Conor
C
Fields
James John
JJ
O'Mahony
Gil U
GU
Lee
D000595:Amino Acid Sequence; D000818:Animals; D001666:Binding Sites, Antibody; D002918:Chymotrypsin; D003001:Cloning, Molecular; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D008280:Magnetics; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013025:Glycine max / Q000737:chemistry*; D014357:Trypsin; D014358:Trypsin Inhibitor, Bowman-Birk Soybean / Q000276:immunology* / Q000302:isolation & purification
D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014358:Trypsin Inhibitor, Bowman-Birk Soybean; D002918:Chymotrypsin; D014357:Trypsin