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Method of measuring available free thyroxine bending sites

  • US 5,244,786 A
  • Filed: 08/13/1990
  • Issued: 09/14/1993
  • Est. Priority Date: 10/02/1987
  • Status: Expired due to Fees
First Claim
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1. A method for directly measuring available thyroxin binding sites in a sample, comprising:

  • (a) combining in an assay medium (1) a β

    -galactosidase substrate, (2) said sample, (3) a β

    -galactosidase acceptor fragment having substantially the same amino acid sequence as a C-terminus region of β

    -galactosidase, and (4) a β

    -galactosidase enzyme donor conjugate comprising a β

    -galactosidase enzyme donor fragment, wherein said donor fragment has substantially the same amino acid sequence as an N-terminus region of β

    -galactosidase complementary to said C-terminus region, linked to a moiety that competes with thyroxin for free thyroxin binding sites, with the proviso that said β

    -galactosidase donor conjugate and said β

    -galactosidase acceptor fragment are not combined in the absence of said sample, and wherein said donor and acceptor fragments form an active β

    -galactosidase enzyme upon complexation, said complexation being modulated if said enzyme donor conjugate binds to a protein having a thyroxin binding site; and

    (b) determining the amount of β

    -galactosidase activity in said assay medium as compared to an assay medium containing a known amount of available thyroxin binding sites.

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