Anne M Bates

US Patent:

50533274, Oct 1, 1991

Filed:

Sep 20, 1988

Appl. No.:

7/246842

Inventors:

David L. Brandon - Berkeley CA
Anne H. Bates - Lafayette CA
Mendel Friedman - Moraga CA

Assignee:

The United States of America as represented by the Secretary of
Agriculture - Washington DC

International Classification:

G01N 3353

US Classification:

435 792

Abstract:

Hybrid cell lines (hybridomas) which produce and secrete high affinity monoclonal antibodies specific for Bowman-Birk inhibitor (BBI) are described. High affinity antibodies to BBI are described that have one or more of the following additional characteristics: (1) they are specific to the active form of BBI, that is, they react and bind with undenatured BBI, but do not bind with BBI which has been denatured by heat or disulfide exchange; (2) they do not react and bind with KTI; (3) they distinguish classical BBI from other BBI's including lima bean protease inhibitor; and (4) they bind BBI-protease complex, e. g. , BBI-chymotrypsin. Immunoassay methods using the monoclonal antibodies to analyze BBI specifically in plant, animal or human tissue or fluid or foodstuffs and techniques for immunoaffinity binding of BBI are described.

US Patent:

54590442, Oct 17, 1995

Filed:

Jul 22, 1991

Appl. No.:

7/733795

Inventors:

David L. Brandon - Berkeley CA
Anne H. Bates - Lafayette CA
Mendel Friedman - Moraga CA

Assignee:

The United States of America as represented by the Secretary of
Agriculture - Washington DC

International Classification:

C07K 1600
C12P 2108

US Classification:

435 792

Abstract:

Hybrid cell lines (hybridomas) which produce and secrete high affinity monoclonal antibodies specific for Bowman-Birk inhibitor (BBI) are described. High affinity antibodies to BBI are described that have one or more of the following additional characteristics: (1) they are specific to the active form of BBI, that is, they react and bind with undenatured BBI, but do not bind with BBI which has been denatured by heat or disulfide exchange; (2) they do not react and bind with KTI; (3) they distinguish classical BBI from other BBI's including lima bean protease inhibitor; and (4) they bind BBI-protease complex, e. g. , BBI-chymotrypsin. Immunoassay methods using the monoclonal antibodies to analyze BBI specifically in plant, animal or human tissue or fluid or foodstuffs and techniques for immunoaffinity binding of BBI are described.

US Patent:

49593102, Sep 25, 1990

Filed:

Sep 28, 1987

Appl. No.:

7/101918

Inventors:

David L. Brandon - Berkeley CA
Anne H. Bates - LaFayette CA
Mendel Friedman - Moraga CA

Assignee:

The United States of America as represented by the Secretary of
Agriculture - Washington DC

International Classification:

C12P 2108
C12N 518
G01N 33577
A61K 39395

US Classification:

435 7

Abstract:

Hybrid cell lines (hybridomas) which produce and secrete monoclonal antibodies with three distinct patterns of recognition are described. The first specificity pattern is defined by antibodies which (1) recognize Kunitz trypsin inhibitor (KTI), one of the principal protease inhibitors found in soybeans, but do not detect the Bowman-Birk inhibitors (BBI), the other major class of protease inhibitors in soybeans; (2) bind to native KTI isoforms a and b but do not react with KTI isoforms a and b which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; and (3) do not recognize native KTI isoform c. The second specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind native KTI isoforms a and c, but do not bind strongly to KTI isoforms a and c which have been denatured by moist heat or alkaline treatment or which have been subjected to disulfide exchange; (3) do not bind KTI isoform b; and (4) bind only weakly to KTI when KTI is complexed with trypsin or a similar enzyme. The third specificity pattern is defined by antibodies that (1) recognize KTI, but do not bind BBI; (2) bind equivalently to native KTI isoforms a, b, and c, but do not bind to KTI which has been denatured by moist heat or alkaline treatment or which has been subjected to disulfide exchange; and (3) bind equivalently to uncomplexed KTI and KTI complexed with trypsin or a similar enzyme. Immunoassay methods using the monoclonal antibodies to analyze native KTI specifically in soy-derived foodstuffs and in tissues of soybean plants, to determine isoform content of a sample, and to determine the amount of KTI complexed with trypsin in a sample are described.