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Enzyme-amplified lanthanide chelate luminescence

  • US 5,262,299 A
  • Filed: 11/13/1990
  • Issued: 11/16/1993
  • Est. Priority Date: 12/04/1989
  • Status: Expired due to Term
First Claim
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1. In a method for detecting luminescence emitted from an excited lanthanide metal chelate associated with an analyte, including the steps of (1) enzymatically converting a selected substrate in a solution by use of an enzyme into a chelator which in the presence of a lanthanide metal ion forms a luminescent lanthanide metal chelate, said substrate in its unconverted form being non-competitive with said lanthanide metal chelate when exposed to radiation which causes said chelate to luminesce and (2) adding said lanthanide metal to said converted substrate to form said lanthanide metal chelate, the improvement comprising:

  • selecting said substrate from the group of salicyl phosphates represented by the formula (I);

    ##STR24## wherein X1 is hydrogen, halogen, C1 to C20 alkyl, C1 to C20 alkyl containing at least one heteroatom, C1 to C20 halogenated alkyl, C5 to C20 halogenated aryl, C5 to C20 aryl, C1 to C20 aryl containing at least one heteroatom, a heteroaromatic group of pyridine, furan, pyrrole, or thiophene which is ring unsubstituted or ring substituted with C1 to C20 alkyl or halogen, trityl, 2, 4-difluorophenyl; and

    X1 is one or more substituents at the 3, 4, 5, or 6 carbon atom of the benzene ring; and

    increasing the pH of said solution containing said enzymatically converted selected salicyl phosphate of formula (I), to a more alkaline pH in forming said lanthanide metal chelate.

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