Generation of monoclonal antibody CIBCgp185 against C-erbB-2 oncoprotein and its clinical evaluation.
The C-erbB-2 proto-oncogene encodes the production of a cell surface receptor protein, with tyrosine kinase activity. Over expression of this gene either due to gene amplification and/or increased transcription has been observed and has been correlated with poor prognosis in patients with Breast (10-33%) and ovarian (20-33%) cancers. The very low levels of expression of C-erbB-2 by normal tissues makes this receptor a potential target for diagnosis and therapy with Monoclonal antibodies raised against its extracellular domain. One such monoclonal antibody designated as CIBCgp185 of IgG2a isotype has been generated in our laboratory using BT474 breast carcinoma cell line as immunogen. This monoclonal antibody immunoprecipitated a 185 KD glycoprotein. The specificity of this antibody was confirmed by the formation of a single discrete band and positive reaction with BT474 antigen in Western blot and Dot blot respectively. Flowcytometric analysis performed using various cancer cell lines revealed that this Monoclonal antibody exhibited high binding affinity with BT474 and SKBR3 cells whichoverexpresses C-erbB-2. By immunoperoxidase test, this antibody stained specifically the tumor cell membrane in frozen tissue sections of breast and ovarian tumors indicating overexpression of the C-erbB-2 product. All these results well correlated with those obtained using a control antibody ICR12, an anti-C-erbB-2 antibody. These studies clearly indicate that Monoclonal antibody CIBCgp185 might prove useful to identify tumors with over expression of C-erbB-2 which are often associated with poor prognosis and early recurrence.
Meenakshi
A
A
Department of Biochemical Oncology, Cancer Institute (WIA), Chennai-600020, India. caninst@md2.vsnl.net.in
Kumar
R S
RS
Kumar
N S
NS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D000090063:Proto-Oncogene Mas; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis / Q000276:immunology* / Q000502:physiology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000090063:Proto-Oncogene Mas; D018719:Erb-b2 Receptor Tyrosine Kinases; C000715470:MAS1 protein, human