Receptor assays of biologically active compounds employing biologically specific receptors

• US 3,901,654 A
• Filed: 06/21/1971
• Issued: 08/26/1975
• Est. Priority Date: 06/21/1971
• Status: Expired due to Term

• Current Assignee: Stanley J. Gross
• Original Assignee: Stanley J. Gross
• Inventors: Gross, Stanley J.
• Primary Examiner(s): Scovronek, Joseph
• Application Number:
  US05/155,261
• Time in Patent Office: 1,527 Days
• Field of Search: 23/23R,23B,253TP
• US Class Current: 436/172
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: Y10T 436/142222 Hetero-O [e.g., ascorbic acid, etc.]
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/200833 Carbonyl, ether, aldehyde or ketone containing
• CPC Class Codes: Y10T 436/203332 Hydroxyl containing

Abstract

This invention relates to novel assays for biologically active substrates employing purified target receptors. The assays comprise fluorescent assays for quenching and enhancement as well as radiochemical assays, employing labeled receptor in one embodiment and labeled substrate in another. Methods for the preparation of these purified receptors are also disclosed.

Claims

1. AFLUORESCENT METHOD OF ASSAYING QUANTITIVELY FOR A COMPOUND WHICH IS SPECIFIC FOR A RECEPTOR COMPOUND, WHICH COMPRISES PRODUCING A MIXTURE, SUITABL FOR FLUOROMETRIC ASSAY, OF (1) A SAMPLE TO BE ANALYZED, (2) A KNOWN QUANTITY OF THE RECEPTOR COMPOUND IN ITS PURIFIED FORM AND WHICH IS BIOLOGICALLY SPECIFIC FOR THE COMPOUND TO BE ASSAYED, AND (3) A KNOWN QUANTITY OF A FLUORESCENCE QUENCHING COMPOUND, POSSESSING AN ABSORBANCE SPECTRUM CORRESPONDING TO THE EMISSION SPECTRUM OF THE RECEPTOR, AND WHICH IS HOMOLOGOUS TO SAID ASSAY COMPOUND SO AS TO BIND WITH SAID RECEPTOR COMPETITIVELY, AN ANALYZING SAID MIXTURE FOR RECEPTOR FLUORESCENCE QUENCHING.

2. The method of claim 1 wherein said fluorescence quenching compound is a fluorescent compound capable of being enhanced or polarized and which is homologous to said assay compound so as to bind with said receptor competitively;

and analyzing said mixture for fluorescence enhancement or polarization of said fluorescent compound.