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The use of anti-beta 1 integrin monoclonal antibody in lung cancer treatment has proven beneficial.Here,we developed a novel monoclonal antibody(mAb),called P5,by immunizing mice with human peripheral blood mononuclear cells(PBMC).Its anti-tumor effect is now being tested,in a clinical phase HI trial,in combinatorial treatments with various chemical drugs.To confirm that P5 indeed binds to beta 1 integrin,cell lysates were immunoprecipitated with commercial anti-beta 1 integrin mAb(TS2/16) and immunoblotted against P5 to reveal a140 kDa molecular weight band,as expected.Immunoprecipitation with P5 followed by LC/MS protein sequence analysis further verified P5 antigen to be beta 1 integrin.Cisplatin treatment upregulated cell surface expression of beta 1 integrin in A549 cells,while causing inhibition of cell growth.When cells were co-treated with different concentrations of P5 mAb,the cisplatin-mediated inhibitory effect was enhanced in a dose-dependent manner.Our findings show that a combinatorial treatment of P5 mAb and cisplatin in A549 cells resulted in a 30%increase in apoptosis,compared to baseline,and significantly more when compared to either the cisplatin or P5 alone group.The entire peptide sequences in CDR from variable region of Ig heavy and light chain gene for P5 mAb are also disclosed.Together,these results provide evidence of the beneficial effect of P5 mAb in combinatorial treatment of human lung adenocarcinoma.
The use of anti-beta 1 integrin monoclonal antibody in lung cancer treatment has proven beneficial. Here, we developed a novel monoclonal antibody (mAb), called P5, by immunizing mice with human peripheral blood mononuclear cells (PBMC) .Its anti-tumor effect is now being tested, in a clinical phase HI trial, in combinatorial treatments with various chemical drugs. To confirm that P5 indeed binds to beta 1 integrin, cell lysates were immunoprecipitated with commercial anti-beta 1 integrin mAb (TS2 / 16) and immunoblotted against P5 to reveal a 140 kDa molecular weight band, as expected. Immunoprecipitation with P5 followed by LC / MS protein sequence analysis further verified P5 antigen to be beta 1 integrin. Disatin treatment upregulated cell surface expression of beta 1 integrin in A549 cells, while causing inhibition of cell growth .When cells were co-treated with different concentrations of P5 mAb, the cisplatin-mediated inhibitory effect was enhanced in a dose-dependent manner. Our findings show that a combinatorial treatment of P5 mAb and cisplatin in A549 cells resulted in a 30% increase in apoptosis, compared to baseline, and significantly more when compared to either the cisplatin or P5 alone group. The entire peptide sequences in CDR from variable region of Ig heavy and light chain gene for P5 mAb are also disclosed in.Together, these results provide evidence of the beneficial effect of P5 mAb in combinatorial treatment of human lung adenocarcinoma.