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Analytical method

  • US 6,984,494 B2
  • Filed: 08/01/2001
  • Issued: 01/10/2006
  • Est. Priority Date: 08/15/2000
  • Status: Active Grant
First Claim
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1. A method for determining the total amount of an anti-HER2 antibody in a fluid sample in the presence of HER2 extracellular domain (ECD), comprising:

  • (a) providing a solid surface dual-coated with first antibody recognizing said anti-HER2 antibody in a free form and a second antibody recognizing said HER2 ECD when bound to said anti-HER2 antibody;

    (b) contacting said dual-coated surface with said fluid sample, whereby said first antibody binds said free anti-HER2 antibody and said second antibody binds said HER2 ECD bound to said anti-HER2 antibody; and

    (c) determining the total amount of said free anti-HER2 antibody and said anti-HER2 antibody bound to said HER2 ECD by contacting the product of step (b) with a detectably labeled secondary antibody recognizing said anti-HER2 antibody, wherein said detectably labeled secondary antibody recognizes an epitope which is different from the epitope recognized by said first antibody and is also different from the epitope recognized by said HER2 ECD, and wherein the total amount of said free anti-HER2 antibody and said anti-HER2 antibody bound to said HER2 ECD is compared with a standard curve generated by using various concentrations of said purified anti-HER2 antibody in place of said fluid sample in step (b).

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