Diagnostic methods and probes
First Claim
1. A method for detecting an analyte in a sample, said method comprising the steps of:
- contacting said sample with a detector probe and a capture probe to form a detector probe-analyte-capture probe complex, wherein said detector probe comprises a moiety having a predetermined pI and comprising a detectable label;
separating said complex from any detector probe that is not bound in said complex;
releasing said moiety from said complex;
isolating said moiety released from said complex and concentrating said moiety by isoelectric focusing; and
detecting said moiety as a measure of the presence of said analyte in said sample.
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Abstract
The invention features methods for detecting a target analyte in a sample. In these methods, a sample is contacted with a detector probe and a capture probe to form a detector probe-analyte-capture probe complex. The detector probe includes a new moiety, having a predetermined pI and containing a detectable label. The complex is isolated from detector probe that is not bound in the complex, the moiety is released from the complex, and the released moiety is concentrated, e.g., by isoelectric focusing or other methods. Detection of the moiety, e.g., at a position in a pH gradient corresponding to its pI, can be used as a measure of the presence and concentration of the analyte in the sample.
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Citations
32 Claims
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1. A method for detecting an analyte in a sample, said method comprising the steps of:
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contacting said sample with a detector probe and a capture probe to form a detector probe-analyte-capture probe complex, wherein said detector probe comprises a moiety having a predetermined pI and comprising a detectable label; separating said complex from any detector probe that is not bound in said complex; releasing said moiety from said complex; isolating said moiety released from said complex and concentrating said moiety by isoelectric focusing; and detecting said moiety as a measure of the presence of said analyte in said sample. - 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)
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32. A method for detecting the presence of a chromosome comprising a chromosomal translocation in a sample, wherein said chromosome comprises a portion of a first chromosome and a portion of a second chromosome, said method comprising the steps of:
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contacting said sample with a capture probe and a detector probe to form a detector probe-analyte-capture probe complex, wherein said capture probe binds to said portion of said first chromosome, or a corresponding RNA transcript thereof, and comprises the first member of a specific binding pair, and said detector probe binds to said portion of said second chromosome, or a corresponding RNA transcript thereof, and comprises a moiety having a predetermined pI and a detectable label; separating said complex from any detector probe that is not bound in said complex by contacting said complex with the second member of said specific binding pair, said second member being immobilized on a solid support; releasing said moiety from said complex; concentrating said moiety by isoelectric focusing; and detecting the concentrated moiety as a measure of the presence of said chromosome containing said chromosomal translocation in said sample.
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Specification