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Analysis of circulating tumor cells, fragments, and debris

  • US 8,329,422 B2
  • Filed: 11/02/2010
  • Issued: 12/11/2012
  • Est. Priority Date: 08/23/2001
  • Status: Expired due to Fees
First Claim
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1. A method for diagnosing disease in a test subject comprising:

  • a. obtaining a biological specimen from a test subject, said specimen comprising a mixed cell population suspected of containing intact rare cells and further comprising;

    i. circulating cell fragments derived from rare cells, orii. circulating cellular debris derived from rare cells;

    b. preparing a magnetically-labeled sample wherein said biological sample is mixed with colloidal magnetic nanoparticles, having sizes sufficiently small so as to have a magnetic moment that ensures complete separation from the specimen when exposed to an external magnet, coupled to a first biospecific ligand which reacts and specifically binds to a common specific binding site present in said intact rare cells, and said cell fragments or said cellular debris, to the substantial exclusion of other specimen components;

    c. contacting said magnetically-labeled sample with at least one additional biospecific ligand conjugated to a label and specifically binding to another specific binding site common amongst said intact rare cells, and said cell fragments or said cellular debris which are bound to the first biospecific ligand in step b, to the substantial exclusion of other specimen components;

    d. subjecting said magnetically-labeled sample to the externally-applied high gradient magnetic field to produce a separated magnetically-labeled fraction which is enriched for said intact rare cells, and said cell fragments or said cellular debris; and

    e. analyzing said labeled rare cells, and said labeled cell fragments or said labeled cellular debris from step d, the presence of said labeled rare cells, said labeled cell fragments, and said labeled cellular debris indicating the presence of disease.

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