Identification of a novel monoclonal antibody recognizing CD133

J Immunol Methods. 2010 Sep 30;361(1-2):110-5. doi: 10.1016/j.jim.2010.07.007. Epub 2010 Jul 30.

Abstract

Human CD133 (prominin-1), a cell surface glycoprotein, is used as a marker of hematopoietic and neural stem cells. Antibodies that recognize a glycosylation-dependent CD133 epitope have been extensively used for enrichment of tumor initiating cells in a variety of cancers. These currently available antibodies are restricted for use in only a subset of biological assays. We have generated a novel anti-human CD133 monoclonal antibody, using a recombinant protein consisting of highly immunogenic amino acid residues selected from the native CD133 protein as an immunogen. The antibody (identified as clone 7) specifically recognizes the CD133 protein in a variety of immunological applications including Western blot, immunofluorescence, flow cytometry and immunohistochemistry. Further, clone 7 specifically recognizes an unmodified CD133 extracellular domain, and not its glycosylated epitope. In conclusion, the specificity and usefulness in a wide range of applications suggest that clone 7 could be a valuable tool to identify CD133 positive cells as well as to target them for therapy.

MeSH terms

  • AC133 Antigen
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / immunology
  • Antigens, CD / immunology*
  • Blotting, Western
  • Caco-2 Cells
  • Epitopes / immunology*
  • Flow Cytometry
  • Glycoproteins / immunology*
  • Humans
  • Immunohistochemistry
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Peptides / immunology*
  • Transfection

Substances

  • AC133 Antigen
  • Antibodies, Monoclonal
  • Antigens, CD
  • Epitopes
  • Glycoproteins
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse