Abstract
CD133, also known as prominin-1, was first described as a cell surface marker on early progenitor and hematopoietic stem cells. It is a five-domain transmembrane protein composed of an N-terminal extracellular tail, two small cytoplasmic loops, two large extracellular loops containing seven potential glycosylation sites, and a short C-terminal intracellular tail. CD133 has been used as a marker to identify cancer stem cells derived from primary solid tumors and as a prognostic marker of gliomas. Herein, we developed novel anti-CD133 monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. We expressed the full length of CD133 in LN229 glioblastoma cells, immunized mice with LN229/CD133 cells, and performed the first screening using flow cytometry. After limiting dilution, we established 100 anti-CD133 mAbs, reacting with LN229/CD133 cells but not with LN229 cells. Subsequently, we performed the second and third screening with Western blot and immunohistochemical analyses, respectively. Among 100 mAbs, 11 strongly reacted with CD133 in Western blot analysis. One of 11 clones, CMab-43 (IgG2a, kappa), showed a sensitive and specific reaction against colon cancer cells, warranting the use of CMab-43 in detecting CD133 in pathological analyses of CD133-expressing cancers.
Highlights
Cancer stem cells (CSCs) share many of the properties of non-neoplastic stem cells
CD133, known as prominin-1, was first described as a cell surface marker on hematopoietic stem cells.[13]. It is a five-transmembrane glycoprotein composed of an N-terminal extracellular tail, two small cytoplasmic loops, two large extracellular loops containing several potential glycosylation sites, and a short C-terminal intracellular tail.[14]. CD133 has been used as a marker to identify CSCs derived from primary solid tumors.[1]. Its expression is used as a prognostic marker of gliomas.[15]
The 11 clones selected by Western blot analysis were further tested, and immunohistochemical analysis demonstrated that 4 clones showed strong staining, 4 showed moderate staining, and 3 showed no staining against colon cancer tissues
Summary
Cancer stem cells (CSCs) share many of the properties of non-neoplastic stem cells. CD133, known as prominin-1, was first described as a cell surface marker on hematopoietic stem cells.[13] It is a five-transmembrane glycoprotein composed of an N-terminal extracellular tail, two small cytoplasmic loops, two large extracellular loops containing several potential glycosylation sites, and a short C-terminal intracellular tail.[14] CD133 has been used as a marker to identify CSCs derived from primary solid tumors.[1] Its expression is used as a prognostic marker of gliomas.[15]. We produced sensitive and specific monoclonal antibodies (mAbs) against CD133, which can be used for flow cytometry, Western blot, and immunohistochemical analysis
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
