Mobile website
Basic Research
Screen for functional monoclonal antibodies and antigens of lung cancer by functional differential cell model
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To screen for functional monoclonal antibodies against lung cancer cell membrane antigens, so as to provide candidate targets and drugs for targeted therapy of lung cancer. Methods:Malignant cells separated from lung carcinoma tissues were used to immunize BALB/c mice. The spleen cells of the immunized mouse were fused with SP2/0 cells to construct functional monoclonal antibody library. Several lung cancer cell lines were domesticated to establish functionaloriented differential sub cell lines of proliferation, invasion and migration, then the antibodies of each clone were screened by immunofluorescence with the functional differential sub cell lines to obtain the functional monoclonal antibodies against cell membranes proteins and then screened by in vitro functional assays. Antigens of functional monoclonal antibodies were identified by Western blotting and the inhibitory effects of individual antibodies were tested by tumor treatment experiments in vivo. Results:Totally 2 893 monoclonal antibody clones were obtained by fusion of spleen cells with SP2/0 cells and 309 clones reacted with the lung cancer cell membranes components. Three functionaloriented differential sub cell line models of lung cancer cells, including the proliferation, invasion and migration, were established for primary screening. Twenty clones of monoclonal antibodies significantly suppressed the lung cancer cell proliferation, 10 monoclonal antibodies could inhibit tumor cell invasion in matrigel and 5 inhibited tumor cell migration（P<0.05 or P<001）. Western blotting assay identified 6 target antigens from these functional antibodies above. In vivo assay further demonstrated that 2 of these functional antibodies significantly inhibited lung tumor growth in mice. Conclusion:Combination of functional antibody library and differential functionaloriented cell models can be used to batch screen for functional monoclonal antibodies, which can suppress the malignant behavior of lung cancer cells in vitro and in vivo, providing a new method for screening the targets for the antimalignancy therapy.
Keywords:
Project Supported:
Supported by the Major Basic Research and Development Program (973) of China（No.2009CB521804）; the National High Technology Research and Development Program (863) of China（No.2006AA02Z479）.
Clc Number:
