Multiplex analysis using membrane-bound sensitizers
First Claim
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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:
- providing a lipid membrane comprising one or more membrane-associated analytes and a lipophilic sensitizer anchored therein;
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 to the binding compound 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 characteristcs;
combining the lipid 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 lipophilic sensitizer in the lipid membrane so that an active species is produced that cleaves cleavable linkages between molecular tags and binding compounds that form said complexes 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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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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15 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 lipid membrane comprising one or more membrane-associated analytes and a lipophilic sensitizer anchored therein;
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 to the binding compound 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 characteristcs;
combining the lipid 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 lipophilic sensitizer in the lipid membrane so that an active species is produced that cleaves cleavable linkages between molecular tags and binding compounds that form said complexes 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, 13, 14, 15)
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Specification