Monoclonal antibody CAb-1 heavy and light chain variable region gene of human carcinoma of large intestine, and its uses
A technology of monoclonal antibody and variable region, which is applied in the fields of application, antibody, gene therapy, etc., and can solve problems such as difficult to obtain high-affinity and high-specificity colorectal cancer monoclonal antibody
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[0015] (1) Cloning of light and heavy chain variable region genes of anti-human colorectal cancer monoclonal antibody CAb-1
[0016] The cell line used is a high-affinity, high-specificity mouse-derived anti-human colorectal cancer monoclonal antibody CAb-1 hybridization obtained by the inventors by using fresh colorectal cancer tissue homogenate to immunize mice and using traditional fusion methods tumor cell lines. The secreted antibody does not bind to tumor-associated antigens such as CEA, 17-1A, TAG-72, CEA, A33, etc., and is aimed at a new tumor-associated glycoprotein antigen with a molecular weight of about 200KD. Its secreted antibody molecular subtype is IgG1κ.
[0017] CAb-1 hybridoma cells (5×10 6 ), using guanidine isothiocyanate one-step method to extract total RNA, and take a small amount for quantification by UV spectrophotometer and detection by 1% formaldehyde-denaturing agarose gel electrophoresis. Then Oligo(dT) 15 (Promega Company) was used as a random...
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