US4859611A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Affinity column and process for detection of low molecular weight toxic substances
Assignee: Boston University
•
Filing: 1986-01-21
•
Granted: 1989-08-22
•
Expires: 2006-01-21
Abstract
Abstract An affinity matrix and a method for the detection of low molecular weight compositions such as aflatoxins are provided utilizing specific monoclonal IgM antibody having an affinity constant not less than about 1×10 9 liters per mole. Methods for the preparation and use of such affinity matrices are also given. The detection is rapid, accurate, reproducible, and allows for quantitative recovery of the composition of interest.
Inventors
- 👤 John D. Groopman
- 👤 Gerald N. Wogan
- 👤 Frederick G. Bargoot
- 👤 Christopher Ferrari
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 05:03
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/14; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/569; G01N33/56961; Y; Y10; Y10S; Y10S436/00; Y10S436/815; Y10S436/824; Y10S436/825
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
150 words
A method for detecting toxins in a test sample, said method comprising: preparing a sample believed to contain a toxin; placing said sample on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody specific for said toxin; eluting said column with a solvent to recover a first eluent, whereby said toxin is retained on said column by said monoclonal antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent to recover a second eluent, whereby said toxin is released from said antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light for detection of the presence of said toxin, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery from said column.
Claim 2
Independent Claim
150 words
A method for detecting toxins in a test sample, said method comprising: preparing a sample believed to contain a toxin; placing said sample on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody specific for said toxin; eluting said column with a solvent to recover a first eluent, whereby said toxin is retained on said column by said monoclonal antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent to recover a second eluent, whereby said toxin is released from said antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light for detection of the presence of said toxin, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery from said column.
Claim 3
Dependent Claim
21 words
A method according to claim 1, wherein said releasing agent comprises a not less than 50% solution of an aprotic solvent.
Claim 4
Dependent Claim
18 words
A method according to claim 2, wherein said aprotic solvent is dimethyl sulfoxide, dimethyl formamide or dimethyl acetamide.
Claim 5
Dependent Claim
18 words
A method according to claim 1, wherein said releasing agent comprises a 50-100% solution of a lipophilic solvent.
Claim 6
Dependent Claim
15 words
A method according to claim 4, wherein said solvent comprises 100% methanol or 100% ethanol.
Claim 7
Dependent Claim
37 words
A method according to claim 5, wherein said toxin is aflatoxin B 1 or aflatoxin M 1 and said antibody is specific for said aflatoxins with an affinity of at least about 1×10 8 liters per mole.
Claim 8
Dependent Claim
26 words
A method according to claim 6, wherein said second eluent is exposed to UV light at 365 nm for visual detection of the presence of aflatoxin.
Claim 9
Dependent Claim
24 words
A method according to claim 6, wherein said second eluent is subjected to quantitative analysis for determining the level of aflatoxin in said sample.
Claim 10
Dependent Claim
23 words
A method according to claim 1, wherein said toxin is a mycotoxin having a molecular weight of not more than about 1,000 daltons.
Claim 11
Dependent Claim
31 words
A method according to claim 9, wherein said toxin is a member selected from the group consisting of aflatoxins, fluoranthene, nitropyrene, nitrosopyrene, nitrofluoranthene, nitrochrysene, aminobiphenyl, and their respective conjugates and derivatives.
Claim 12
Dependent Claim
15 words
A method according to claim 1, wherein said sample is serum, urine or food product.
Claim 13
Dependent Claim
16 words
A method according to claim 11, wherein said food product is milk, corn or peanut products.
Claim 14
Independent Claim
146 words
A method for detecting aflatoxins in a test sample, said method comprising: placing a sample believed to contain aflatoxin on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody for said aflatoxin; eluting said column with a solvent to recover a first eluent, whereby said aflatoxin is retained on said column by said monoclonal antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent to recover a second eluent, whereby said aflatoxin is released from said monoclonal antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light for detection of the presence of said aflatoxin, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery from said column.
Claim 15
Independent Claim
146 words
A method for detecting aflatoxins in a test sample, said method comprising: placing a sample believed to contain aflatoxin on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody for said aflatoxin; eluting said column with a solvent to recover a first eluent, whereby said aflatoxin is retained on said column by said monoclonal antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent to recover a second eluent, whereby said aflatoxin is released from said monoclonal antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light for detection of the presence of said aflatoxin, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery from said column.
Claim 16
Dependent Claim
21 words
A method according to claim 13, wherein said releasing agent comprises a not less than 50% solution of an aprotic solvent.
Claim 17
Dependent Claim
18 words
A method according to claim 14, wherein said aprotic solvent is dimethyl sulfoxide, dimethyl formamide or dimethyl acetamide.
Claim 18
Dependent Claim
18 words
A method according to claim 13, wherein said releasing agent comprises a 50-100% solution of a lipophilic solvent.
Claim 19
Dependent Claim
15 words
A method according to claim 16, wherein said solvent comprises 100% methanol or 100% ethanol.
Claim 20
Dependent Claim
26 words
A method according to claim 13, wherein said antibody has an affinity of at least about 1×10 8 liter per mole for the aflatoxin of interest.
Claim 21
Dependent Claim
15 words
A method according to claim 13, wherein said sample comprises serum, urine or food product.
Claim 22
Dependent Claim
16 words
A method according to claim 19, wherein said food product is milk, corn or peanut products.
Claim 23
Dependent Claim
18 words
A method according to claim 13, wherein said second eluent is exposed to UV light at 365 nm.
Claim 24
Dependent Claim
24 words
A method according to claim 13, wherein said second eluent is subjected to quantitative analysis for determining the level of aflatoxin in said sample.
Claim 25
Independent Claim
151 words
A method for detecting aflatoxins in a test sample, said method comprising: placing a sample believed to contain aflatoxin on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody for said aflatoxin; eluting said column with a solvent to recover a first eluent, whereby said aflatoxins are retained on said column by said antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent comprised of ethanol or methanol to recover a second eluent, whereby said aflatoxins are released from said antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light at 365 nm for detection of the presence of aflatoxins, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery form said column.
Claim 26
Independent Claim
151 words
A method for detecting aflatoxins in a test sample, said method comprising: placing a sample believed to contain aflatoxin on an affinity column comprised of a solid phase sorbant material and, immobilized thereon, a monoclonal antibody for said aflatoxin; eluting said column with a solvent to recover a first eluent, whereby said aflatoxins are retained on said column by said antibody and separated from all other ingredients of said sample; eluting said column with a releasing agent comprised of ethanol or methanol to recover a second eluent, whereby said aflatoxins are released from said antibody and recovered in said second eluent; and subjecting said second eluent to fluorescence measurement by exposing said second eluent to UV light at 365 nm for detection of the presence of aflatoxins, with the proviso that said second eluent is not subjected to high pressure liquid chromatography or other purification subsequent to recovery form said column.
Claim 27
Dependent Claim
24 words
A method according to claim 23, wherein said antibody has an affinity of at least about 1×10 8 liters per mole to said aflatoxins.
Claim 28
Dependent Claim
15 words
A method according to claim 23, wherein said sample comprises serum, urine or food product.
Claim 29
Dependent Claim
16 words
A method according to claim 25, wherein said food product is milk, corn or peanut products.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-08-21 | 1993-02-20 | 1993-08-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-08-21 | 1997-02-20 | 1997-08-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-08-21 | 2001-02-20 | 2001-08-22 | $7700.00 | Window Closed / Lapsed |