Patent US5053327A successfully refreshed from Google Patents.
US5053327A
Expired / Lapsed
US
⏱️ 0.0 years remaining
High affinity monoclonal antibodies to bowman-birk inhibitor and immunoassay methods
Assignee: United States Department of Agriculture
•
Filing: 1988-09-20
•
Granted: 1991-10-01
•
Expires: 2008-09-20
Abstract
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.
Inventors
- 👤 David L. Brandon
- 👤 Anne H. Bates
- 👤 Mendel Friedman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 20:49
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/38; Y; Y10; Y10S; Y10S530/00; Y10S530/808; Y10S530/809
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
38 words
A continuous hybrid cell line (ATCC No. HB 9657) which produces and secretes monoclonal antibodies which bind native Bowman-Birk inhibitor (BBI) with an affinity constant of at least 8×10 7 M -1 and do not bind denatured BBI.
Claim 2
Dependent Claim
38 words
A continuous hybrid cell line (ATCC No. HB 9657) which produces and secretes monoclonal antibodies which bind native Bowman-Birk inhibitor (BBI) with an affinity constant of at least 8×10 7 M -1 and do not bind denatured BBI.
Claim 3
Dependent Claim
21 words
The continuous hybrid cell line described in claim 1 wherein said monoclonal antibodies do not bind with Kunitz trypsin inhibitor (KTI).
Claim 4
Dependent Claim
19 words
The continuous hybrid cell line described in claim 1 wherein said monoclonal antibodies are selective for an individual BBI.
Claim 5
Dependent Claim
26 words
The continuous hybrid cell line described in claim 1 wherein said monoclonal antibodies bind BBI which is bound to a protein at its chymotrypsin reactive site.
Claim 6
Dependent Claim
25 words
The continuous hybrid cell line described in claim 1 obtained using an immunization preparation comprising BBI which has not been treated with a crosslinking agent.
Claim 7
Dependent Claim
32 words
The continuous hybrid cell line of claim 1 which produces and secretes high affinity monoclonal antibodies to soybean BBI which do not bind with KTI, denatured BBI, or lima bean inhibitor (LBI).
Claim 8
Dependent Claim
32 words
Monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI.
Claim 9
Dependent Claim
32 words
Monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI.
Claim 10
Dependent Claim
12 words
The monoclonal antibody of claim 7 which does not bind with KTI.
Claim 11
Dependent Claim
13 words
The monoclonal antibody of claim 7 which is selective for an individual BBI.
Claim 12
Dependent Claim
20 words
The monoclonal antibody of claim 7 which binds BBI which is bound to a protein at its chymotrypsin reactive site.
Claim 13
Dependent Claim
15 words
Monoclonal antibody of claim 7 which does not bind with KTI, denatured BBI, or LBI.
Claim 14
Independent Claim
127 words
A method for determining the amount of native BBI in a sample, which comprises: (a) incubating a sample with a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI to obtain a mixture containing monoclonal antibody bound to native BBI in said sample and unbound antibody; (b) contacting said mixture with a BBI-coated solid phase to bind said unbound antibody; (c) separating said solid phase-bound antibody from said contacted mixture; (d) detecting said separated solid phase-bound monoclonal antibody with labeled antibody; and (e) determining the amount of native BBI in said sample by comparing the amount of detected labeled antibody to a standard curve.
Claim 15
Independent Claim
127 words
A method for determining the amount of native BBI in a sample, which comprises: (a) incubating a sample with a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI to obtain a mixture containing monoclonal antibody bound to native BBI in said sample and unbound antibody; (b) contacting said mixture with a BBI-coated solid phase to bind said unbound antibody; (c) separating said solid phase-bound antibody from said contacted mixture; (d) detecting said separated solid phase-bound monoclonal antibody with labeled antibody; and (e) determining the amount of native BBI in said sample by comparing the amount of detected labeled antibody to a standard curve.
Claim 16
Independent Claim
125 words
A method for determining the amount of native BBI in a sample, which comprises: (a) incubating a sample with labeled BBI to obtain a mixture; (b) contacting said mixture with a solid phase coated with monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI to bind native BBI in said sample in said mixture and labeled BBI in said mixture; (c) separating said solid phase-bound labeled BBI from said contacted mixture; (d) detecting said separated solid phase-bound labeled BBI; and (e) determining the amount of native BBI in said sample by comparing the amount of detected labeled BBI to a standard curve.
Claim 17
Independent Claim
125 words
A method for determining the amount of native BBI in a sample, which comprises: (a) incubating a sample with labeled BBI to obtain a mixture; (b) contacting said mixture with a solid phase coated with monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI to bind native BBI in said sample in said mixture and labeled BBI in said mixture; (c) separating said solid phase-bound labeled BBI from said contacted mixture; (d) detecting said separated solid phase-bound labeled BBI; and (e) determining the amount of native BBI in said sample by comparing the amount of detected labeled BBI to a standard curve.
Claim 18
Dependent Claim
40 words
The method of claim 12 wherein said sample is selected from the group consisting of plant, animal or human tissue or fluid, a plant seed, a substance containing the product of a gene derived from a plant, and a foodstuff.
Claim 19
Dependent Claim
43 words
The method of claim 13 wherein said sample is selected from the group consisting of plant, animal or human tissue or fluid, a plant seed, a substance containing the product of a gene derived from a plant, and a foodstuff or plant tissue.
Claim 20
Independent Claim
69 words
A method for attaching BBI to a solid phase, which comprises: (a) attaching a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI, to a solid phase by a covalent or noncovalent bond; and (b) contacting said solid phase with BBI to form solid phase-bound BBI.
Claim 21
Independent Claim
69 words
A method for attaching BBI to a solid phase, which comprises: (a) attaching a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI, to a solid phase by a covalent or noncovalent bond; and (b) contacting said solid phase with BBI to form solid phase-bound BBI.
Claim 22
Dependent Claim
99 words
A kit useful for the detection of native BBI in a sample, which comprises a carrier being compartmentalized to receive one or more containers in close confinement therein; a first container means comprising a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI; and a second container means comprising detection means selected from the group consisting of labeled BBI, labeled monoclonal antibody specific for BBI, and labeled immunoglobulin which binds to monoclonal antibody specific for BBI.
Claim 23
Dependent Claim
99 words
A kit useful for the detection of native BBI in a sample, which comprises a carrier being compartmentalized to receive one or more containers in close confinement therein; a first container means comprising a monoclonal antibody produced by hybridoma cell line ATCC No. HB 9657 which binds native BBI with an affinity constant of at least 8×10 7 M -1 and does not bind denatured BBI; and a second container means comprising detection means selected from the group consisting of labeled BBI, labeled monoclonal antibody specific for BBI, and labeled immunoglobulin which binds to monoclonal antibody specific for BBI.
References Cited (Backward Citations)
US4196265A
Cited
US4271145A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-09-30 | 1995-04-01 | 1995-10-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-09-30 | 1999-04-01 | 1999-10-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-09-30 | 2003-04-01 | 2003-10-01 | $7700.00 | Window Closed / Lapsed |