Multiplex analysis using membrane-bound sensitizers
First Claim
1. A method for determining the presence or absence of one or more membrane-associated analytes, the method comprising the steps of:
- providing a sensitizer-treated membrane comprising the one or more membrane-associated analytes;
providing one or more binding compounds, such that there is at least one binding compound specific for each of the one or more membrane-associated analytes, each binding compound having one or more molecular tags, each molecular tag being attached by a cleavable linkage, and the molecular tags of each binding compound being distinguishable from those of every other binding compound by one or more physical and/or optical characteristics;
combining the sensitizer-treated membrane and the plurality of binding compounds specific for the one or more membrane-associated analytes such that in the presence of a membrane-associated analyte a complex is formed between such membrane-associated analyte and a binding compound specific therefor;
activating the sensitizer-treated membrane so that an active species is produced that cleaves the cleavable linkage of the binding compounds forming such complex so that molecular tags are released; and
separating and identifying the released molecular tags by the one or more physical characteristics to determine the one or more membrane-associated analytes in the sample.
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Accused Products
Abstract
The present invention is directed to methods and compositions for determining the presence, absence, and/or amounts of one or more membrane-associated analytes in a sample. In accordance with the invention, binding compounds derivatized with releasable molecular tags specifically bind to selected membrane-associated analytes, after which the molecular tags are released upon activation of cleavage moieties, or sensitizers, anchored in the same membrane as the membrane-associated analytes. The released molecular tags are then identified by their distinct separation and detection characteristics.
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Citations
20 Claims
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1. A method for determining the presence or absence of one or more membrane-associated analytes, the method comprising the steps of:
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providing a sensitizer-treated membrane comprising the one or more membrane-associated analytes;
providing one or more binding compounds, such that there is at least one binding compound specific for each of the one or more membrane-associated analytes, each binding compound having one or more molecular tags, each molecular tag being attached by a cleavable linkage, and the molecular tags of each binding compound being distinguishable from those of every other binding compound by one or more physical and/or optical characteristics;
combining the sensitizer-treated membrane and the plurality of binding compounds specific for the one or more membrane-associated analytes such that in the presence of a membrane-associated analyte a complex is formed between such membrane-associated analyte and a binding compound specific therefor;
activating the sensitizer-treated membrane so that an active species is produced that cleaves the cleavable linkage of the binding compounds forming such complex so that molecular tags are released; and
separating and identifying the released molecular tags by the one or more physical characteristics to determine the one or more membrane-associated analytes in the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A composition for determining the presence or absence of one or more membrane-associated analytes, the composition comprising:
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a sensitizer-treated membrane containing one or more membrane-associated analytes; and
one or more binding compounds, such that there is at least one binding compound specific for each of the one or more membrane-associated analytes, each binding compound having one or more molecular tags, each molecular tag being attached by a cleavable linkage, and the molecular tags of each binding compound being distinguishable from those of every other binding compound by one or more physical and/or optical characteristics;
wherein at least one binding compound of the plurality is specifically bound to at least one membrane-associated analyte. - View Dependent Claims (14, 15, 16, 17, 18, 19)
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20. A kit for determining the presence or absence of one or more membrane-associated analytes, the kit comprising:
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a lipophilic photosensitizer for generating a photosensitizer-treated membrane having one or more membrane-associated analytes; and
one or more binding compounds, such that there is at least one binding compound specific for each of the one or more membrane-associated analytes, each binding compound having one or more molecular tags, each molecular tag being attached by a cleavable linkage, and the molecular tags of each binding compound being distinguishable from those of every other binding compound by one or more physical and/or optical characteristics.
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Specification