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Biospecific desorption microflow systems and methods for studying biospecific interactions and their modulators

  • US 20040005582A1
  • Filed: 12/19/2002
  • Published: 01/08/2004
  • Est. Priority Date: 08/10/2000
  • Status: Abandoned Application
First Claim
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1. A microfluidic biospecific desorption assay method for characterizing the binding site of a protein, said method comprising:

  • (1) establishing a buffer flow through a microchannel in fluidic contact with an immobilized binding complex comprising a first immobilized binding pair member and a second labeled binding pair member;

    wherein one of the first or second members is the protein or a fragment of the protein; and

    wherein the protein or protein fragment is bound to the other binding pair member via the binding site;

    (2) introducing a polypeptide into the buffer flow;

    wherein the polypeptide has an amino acid subsequence of the protein;

    (3) detecting the desorption of the label following introduction of the polypeptide; and

    repeating steps (2) and (3) for each of a plurality of polypeptides of differing amino acid sequences, wherein at least one of the polypeptides comprises the binding site;

    whereby the polypeptide comprising the binding site is identified and the binding site is thereby localized to a portion of the protein having the amino acid sequence of the polypeptide comprising the binding site.

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