Fluorescent immunoassay employing a phycobiliprotein labeled ligand or receptor

• US 4,520,110 A
• Filed: 12/30/1982
• Issued: 05/28/1985
• Est. Priority Date: 10/06/1981
• Status: Expired due to Term

• Current Assignee: Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Glazer, Alexander N., Stryer, Lubert, Oi, Vernon T.
• Primary Examiner(s): Marantz, Sidney
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US06/454,768
• Time in Patent Office: 880 Days
• Field of Search: 424/3, 424/7, 424/85, 424/89, 436/528, 436/546.7, 436/805, 436/826, 436/500, 436/501, 436/519, 436/536, 436/546, 436/800, 260/112 R, 260/112 B
• US Class Current: 436/501
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 530/825 Bacteria

Abstract

Sensitive detection techniques and compositions for such techniques employing fluorescent proteins having bilin prosthetic groups as labels i.e. phycobiliprotein. The bilin containing proteins can be conjugated to ligands or receptors for use in systems involving ligand-receptor binding for the analysis, detection or separation of ligands and receptors. Particularly, one or more of the bilin containing proteins may be used as labels in conjuction with each other or other fluorescers for defining subsets of naturally occurring aggregations e.g. cells.

Claims

1. In a fluorescent immunoassay employing as a reagent a fluorescent compound conjugated to a member of a specific binding pair where said pair consists of ligand and receptor, and said immunoassay is for the determination of a member, the binding of the conjugate to a member being indicative of the presence of said member, the improvement which comprises employing in said assay a fluorescent compound comprising a phycobiliprotein conjugated to a member of a specific binding pair consisting of ligand and receptor.

2. In a fluorescent method for detecting cells having a first specific determinant site, employing for fluorescent labeling a fluorescent reagent comprising a fluorescer and a member of a first specific binding pair, the improvement which comprises employing a fluorescent compound comprising a phycobiliprotein conjugated to a member of a specific binding pair consisting of ligand and receptor.

3. In a method according to claim 2, wherein a second fluorescent reagent is employed having different absorption and emission characteristics from said fluorescent compound.

4. In a method for detecting the presence of a determinant site or a receptor, by employing a fluorescent reagent having a fluorescer bound to a member of a specific binding pair, wherein the binding of said fluorescent reagent to said determinant site or receptor is determined as indicative of the presence of said determinant site or said receptor, the improvement which comprises, employing a fluorescent compound comprising a phycobiliprotein conjugated to a member of a specific binding pair consisting of ligand and receptor.