title: Identification of a novel monoclonal antibody recognizing CD133.
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.
authors.affiliation: Department of Pharmaceutics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Suresh Kumar
authors.initials: SK
authors.lastname: Swaminathan
authors.forename: Michael R
authors.initials: MR
authors.lastname: Olin
authors.forename: Colleen L
authors.initials: CL
authors.lastname: Forster
authors.forename: Karen S Santa
authors.initials: KS
authors.lastname: Cruz
authors.forename: Jayanth
authors.initials: J
authors.lastname: Panyam
authors.forename: John R
authors.initials: JR
authors.lastname: Ohlfest
mesh_terms: D000071916:AC133 Antigen; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D015153:Blotting, Western; D018938:Caco-2 Cells; D000939:Epitopes / Q000276:immunology*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D014162:Transfection
chemical_list: D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000939:Epitopes; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse