Affinity column and process for detection of low molecular weight toxic substances
First Claim
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1. 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.
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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 1×109 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.
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Citations
26 Claims
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1. A method for detecting toxins in a test sample, said method comprising:
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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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for detecting aflatoxins in a test sample, said method comprising:
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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. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A method for detecting aflatoxins in a test sample, said method comprising:
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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. - View Dependent Claims (24, 25, 26)
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Specification