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Monolayer and electrode for detecting a label-bearing target and method of use thereof

  • US 6,127,127 A
  • Filed: 04/22/1999
  • Issued: 10/03/2000
  • Est. Priority Date: 06/27/1995
  • Status: Expired due to Term
First Claim
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1. A method of determining the presence of a label-bearing target in a sample, comprising:

  • (a) contacting a non-conductive self-assembled monolayer on an electrode having a conductive working surface, said monolayer comprising phosphonate molecules having at the minimum at least one phosphonate group and at least on R1 group covalently bound to a member of a binding pair, and through which monolayer a transition metal complex can freely move from reactants immobilized on the monolayer to the conductive working surface to transfer electrons to the conductive working surface, with a sample suspected of containing a label-bearing target which is capable of being oxidized in an oxidation-reduction reaction, so that the immobilized member of the binding pair and the target, if present, form a target complex on the monolayer;

    (b) contacting the monolayer and the target complex, if present, with a transition metal complex that oxidizes the label-bearing target in an oxidation-reduction reaction between the transition metal complex and the label-bearing target from which label-bearing target there is electron transfer to the transition metal complex, resulting in regeneration of the reduced form of the transition metal complex as part of a catalytic cycle;

    (c) detecting the oxidation-reduction reaction; and

    (d) determining the presence or absence of the target from the detected oxidation-reduction reaction.

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