Compositions and methods for performing assays
AssigneePalo Alto Research Center Incorporated
InventorsMichael I. RECHT, Peter KIESEL, Joerg MARTINI, Francisco E. TORRES
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Abstract
The disclosure relates to compositions for use in assays, the compositions comprising at least one latent fluorophore including at least one enzyme-reactive quenching group and a conjugative group; and a support connectable to the latent fluorophore by the conjugative group. The disclosure further relates to methods of measuring the presence and/or concentration of an analyte, as well as methods of measuring the relative activity of at least two enzymes.
Claims17
A method of detecting and/or measuring the concentration of an analyte in a sample, the method comprising:
- a. providing a composition comprising:
- i. at least one latent fluorophore comprising at least one enzyme-reactive quenching group and at least one conjugative group, and
- ii. at least one support connectable to the at least one latent fluorophore by the at least one conjugative group;
- b. providing a test sample and a reference sample;
- c. providing at least one first unquenching enzyme capable of releasing the enzyme-reactive quenching group from the at least one latent fluorophore, and at least one second enzyme capable of reacting with at least one analyte;
- d. contacting the test sample with the composition, the at least one first unquenching enzyme, and the at least one second enzyme;
- and
- e. contacting the reference sample with the composition and the at least one first unquenching enzyme.
A method of measuring the activities of at least two enzymes in a sample, the method comprising:
- a. providing a first fluorophore composition comprising:
- i. at least one first enzyme-reactive latent fluorophore comprising at least one first enzyme-reactive quenching group and at least one first conjugative group, and
- ii. at least one support connectable to the at least one first latent fluorophore by at least one first conjugative group;
- b. providing a second fluorophore composition comprising:
- i. at least one second enzyme-reactive latent fluorophore comprising at least one second enzyme-reactive quenching group and at least one second conjugative group, wherein the at least one second enzyme-reactive latent fluorophore is different from said first enzyme-reactive latent fluorophore in said first fluorophore composition, and
- ii. at least one support connectable to the at least one first latent fluorophore by at least one second conjugative group;
- c. providing a test sample to be analyzed and a reference sample to be analyzed;
- d. providing at least one first unquenching enzyme capable of releasing the enzyme-reactive quenching group from the first enzyme-reactive latent fluorophore, and at least one second unquenching enzyme capable of releasing the enzyme-reactive quenching group from the second enzyme-reactive latent fluorophore;
- e. contacting the test sample with the first and second latent fluorophore compositions, the at least one first unquenching enzyme, and the at least one second unquenching enzyme;
- and
- f. contacting the reference sample with the first and second latent fluorophore compositions.
Dependencies are inferred from the claim text.