Analyte detection in particulate-containing samples
AssigneeCytosignet, Inc.
InventorNathan L. Smith, III
Life of this patent
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Abstract
A method of determining the presence and quantity of an analyte of interest in a particulate-containing sample is disclosed, as is a construct for use in the method. The method is particularly useful for determining an analyte in whole blood and in fermentation suspensions. The construct is comprised of a first moiety, which is a particulate-binding moiety and a second moiety, which binds the analyte of interest.
Claims40
A method of determining an analyte of interest in a semen sample, comprising the steps of:
- a) contacting the semen sample with a construct comprised of a first moiety which binds to sperm cells present in the sample as obtained and a second moiety which binds to the analyte of interest, under conditions appropriate for binding of sperm cells to the first moiety of the construct and binding of analyte of interest to the second moiety of the construct;
- and
- b) detecting agglutination of sperm cells in the sample after contacting the sample with the construct of step (a).
A composition for detecting endotoxin in whole blood, comprising a construct consisting essentially of two moieties joined together:
- a first moiety which is a lectin which binds red blood cells present in the sample as obtained, but not to the analyte of interest and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating red blood cells in the sample in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A method of determining endotoxin in a particulate-containing sample, comprising the steps of:
- a) contacting the sample with a construct comprised of a first moiety which is a lectin having binding specificity for particulates present in the sample as obtained, but not for endotoxin, and a second moiety which is polymyxin B, under conditions appropriate for binding of particulate to the first moiety of the construct and binding of endotoxin to the second moiety of the construct, wherein binding of the particulate in the sample to the lectin results in agglutination and the presence of endotoxin in the sample inhibits agglutination;
- and
- b) detecting agglutination of particulates in the sample after contacting the sample with the construct of step (a),
- whereby inhibition of agglutination is observed in the presence of endotoxin.
A method of determining endotoxin in an equine whole blood sample, comprising the steps of:
- a) contacting the whole blood sample with a construct comprised of a first moiety which is a lectin which binds specifically to red blood cells present in the whole blood sample as obtained, but not to endotoxin, and a second moiety which is polymyxin B, under conditions appropriate for binding of red blood cells to the first moiety of the construct and binding of endotoxin to the second moiety of the construct;
- and
- b) determining the extent to which agglutination of the whole blood sample occurs, wherein inhibition of agglutination is indicative of the presence of endotoxin in the whole blood sample.
A method of determining endotoxin in a semen sample, comprising the steps of:
- a) contacting the semen sample with a construct comprised of a first moiety which binds to sperm cells present in the sample as obtained and a second moiety which is polymyxin B, under conditions appropriate for binding of sperm cells to the first moiety of the construct and binding of endotoxin to the second moiety of the construct, wherein binding of sperm cells in the sample to the first moiety results in agglutination of sperm cells and the presence of endotoxin in the sample inhibits agglutination of sperm cells;
- and
- b) detecting agglutination of sperm cells in the sample after contacting the sample with the construct of step (a),
- whereby inhibition of agglutination is observed in the presence of endotoxin.
A construct for detecting endotoxin in a whole blood sample, consisting essentially of two moieties joined together:
- a first moiety which is a lectin which binds red blood cells, but not endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating red blood cells in the sample in the absence of endotoxin, and wherein the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time following addition of the construct to the sample as compared to an appropriately-selected control.
A composition for determining endotoxin in a whole blood sample, comprising a construct comprising two moieties joined together:
- a first moiety which is a lectin which binds red blood cells present in the whole blood sample as obtained, but not to endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating red blood cells in the sample in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A kit for determining endotoxin in a whole blood sample comprising a construct and an appropriate solvent, the construct comprising two moieties joined together:
- a first moiety which is a lectin which binds to red blood cells, but not to the analyte of interest, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating red blood cells in the sample in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A construct for determining endotoxin in semen, comprising two moieties joined together:
- a first moiety which is a lectin which binds to selected particulates present in semen as obtained, but not to endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating the selected particulates present in the sample in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A construct for detecting endotoxin in a semen sample, comprising a first moiety which is a lectin which binds to sperm cells, but not to endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating sperm in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of the endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A kit for detecting endotoxin in a semen sample comprising a construct and an appropriate solvent, said construct comprising a first moiety which is a lectin which binds to sperm cells, but not to endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating sperm in the absence of endotoxin, and wherein, following addition of the construct to the sample, the presence of endotoxin in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
A method of detecting endotoxin in a whole blood sample, comprising the steps of:
- a) contacting the whole blood sample with a construct comprised of a first moiety which is a lectin and which binds specifically to red blood cells present in the sample as obtained, and a second moiety which is polymyxin B, under conditions appropriate for binding of red blood cells to the first moiety of the construct and binding of endotoxin to the second moiety, wherein binding of red blood cells in the sample to the first moiety results in agglutination of red blood cells and the presence of endotoxin in the sample inhibits agglutination of red blood cells;
- and
- b) detecting agglutination of the whole blood sample, whereby inhibition of agglutination is observed in the presence of endotoxin.
A construct for detecting endotoxin in a whole blood sample comprising two moieties joined together:
- a first moiety which is a lectin and which binds specifically to red blood cells present in the sample as obtained, but not to endotoxin, and a second moiety which is polymyxin B, wherein the construct is capable of agglutinating red blood cells in the sample in the absence of endotoxin, and wherein, following addition of construct to the sample, the presence of the analyte in the sample leads to a decrease in the time required for agglutination or in the extent of agglutination at a selected time as compared to an appropriately-selected control.
Dependencies are inferred from the claim text.