Photoprotein conjugates and methods of use thereof

• US 5,486,455 A
• Filed: 08/22/1994
• Issued: 01/23/1996
• Est. Priority Date: 02/12/1993
• Status: Expired due to Term

• Current Assignee: The University of Georgia Research Foundation, Inc.
• Original Assignee: Sealite Sciences, Inc.
• Inventors: Stults, Nancy L.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/293,648
• Time in Patent Office: 519 Days
• Field of Search: 435/6, 435/5, 435/7.1, 536/26.6
• US Class Current: 435/6.16
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

The present invention encompasses a method of synthesis of conjugates of photoproteins that retain all or a substantial portion of the luminescent activity of underivatized photoprotein. According to the present invention photoproteins may be conjugated with a variety of binding including streptavidin/avidin, glyco-proteins, lectins, hormones, antigens, drugs, antibodies and antigen binding fragments thereof, or any other selectively bindable reagent by chemical crosslinking means. The present invention also encompasses conjugates produced by this method, and methods of use of such conjugates.

Claims

1. A photoprotein-binding reagent conjugate composition comprising,

a sulfhydryl-activated aequorin coupled to a sulfhydryl-reactive binding reagent capable of binding specifically to an analyte, to form a conjugate capable of producing light in a luminescent reaction, wherein said aequorin is sulfhydryl-activated by 2-imithiolane (Traut'"'"'s reagent), and said binding reagent is made sulfhydryl-reactive by succinimidyl 4-(N-maleimidomethyl)cyclohexane, (SMCC).