Detecting receptor oligomerization
First Claim
1. A method for detecting a dimer in a sample, the method comprising:
- mixing (i) a sample, which contains a dimer comprising a first membrane-associated analyte and a second membrane-associated analyte;
(ii) a cleaving probe, which is capable of binding the first membrane-associated analyte and has a cleavage-inducing moiety with an effective proximity; and
(iii) one or more binding compounds, at least one of which is capable of binding the second membrane-associated analyte, and each binding compound has one or more molecular tags attached thereto by a cleavable linkage, wherein cleavage of the cleavable linkage(s) within the effective proximity of the cleaving-inducing moiety of the cleaving probe releases the molecular tag(s);
wherein detecting the released molecular tag(s) detects the dimer.
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Abstract
Methods are provided for detecting formation of oligomeric complexes of molecules on the surface of cell membranes. These methods employ pairs of tagged probes and cleaving probes, each of which binds specificly to a cell surface molecule. The tagged probe includes a molecular tag that is linked to a first binding compound through a cleavable linkage, and the cleaving probe includes a second binding agent and a cleavage-inducing moiety that can cleave the linkage when within a defined proximity thereto. Binding of the two probes to cell surface molecules that have formed an oligomeric complex results in release of the molecular tag from the binding compound, providing a measure of formation of the complex.
87 Citations
31 Claims
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1. A method for detecting a dimer in a sample, the method comprising:
- mixing (i) a sample, which contains a dimer comprising a first membrane-associated analyte and a second membrane-associated analyte;
(ii) a cleaving probe, which is capable of binding the first membrane-associated analyte and has a cleavage-inducing moiety with an effective proximity; and
(iii) one or more binding compounds, at least one of which is capable of binding the second membrane-associated analyte, and each binding compound has one or more molecular tags attached thereto by a cleavable linkage, wherein cleavage of the cleavable linkage(s) within the effective proximity of the cleaving-inducing moiety of the cleaving probe releases the molecular tag(s);wherein detecting the released molecular tag(s) detects the dimer. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
- mixing (i) a sample, which contains a dimer comprising a first membrane-associated analyte and a second membrane-associated analyte;
Specification