Department of Pharmaceutics, University of Minnesota, Minneapolis, MN 55455, USA.
J Immunol Methods. 2010 Sep 30;361(1-2):110-5. doi: 10.1016/j.jim.2010.07.007. Epub 2010 Jul 30.
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.
人 CD133（prominin-1）是一种细胞表面糖蛋白，被用作造血和神经干细胞的标志物。识别糖基化依赖性 CD133 表位的抗体已被广泛用于多种癌症中肿瘤起始细胞的富集。这些现有的抗体仅限于在生物测定的一部分中使用。我们使用一种重组蛋白作为免疫原，该重组蛋白由从天然 CD133 蛋白中选择的高度免疫原性氨基酸残基组成，产生了一种新型抗人 CD133 单克隆抗体。该抗体（鉴定为克隆 7）在包括 Western blot、免疫荧光、流式细胞术和免疫组织化学在内的各种免疫应用中特异性识别 CD133 蛋白。此外，克隆 7 特异性识别未经修饰的 CD133 细胞外结构域，而不是其糖基化表位。总之，其在广泛应用中的特异性和有用性表明，克隆 7 可能是一种识别 CD133 阳性细胞并将其作为治疗靶点的有价值的工具。
