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Establishment of CMab-43, a Sensitive and Specific Anti-CD133 Monoclonal Antibody, for Immunohistochemistry
- Source :
- Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
- Publication Year :
- 2017
- Publisher :
- Mary Ann Liebert, Inc., publishers, 2017.
-
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.
- Subjects :
- 0301 basic medicine
medicine.drug_class
Immunology
Biology
Monoclonal antibody
Flow cytometry
03 medical and health sciences
Epitopes
Mice
0302 clinical medicine
Western blot
Cancer stem cell
Cell Line, Tumor
medicine
Extracellular
Immunology and Allergy
Animals
Humans
CD133
AC133 Antigen
neoplasms
medicine.diagnostic_test
Antibodies, Monoclonal
Original Articles
Glioma
Molecular biology
Immunohistochemistry
carbohydrates (lipids)
Haematopoiesis
030104 developmental biology
colon cancer
monoclonal antibody
030220 oncology & carcinogenesis
embryonic structures
Stem cell
Peptides
Subjects
Details
- Language :
- English
- ISSN :
- 21679436
- Volume :
- 36
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
- Accession number :
- edsair.doi.dedup.....672aacf61d38faca89260bbdb51a5c13