采用抗体库技术筛选抑制肺癌的功能性抗体

Identification of Functional Antibodies of Lung Cancer by Screening of Functional Antibody Library

目的研制抑制肺癌细胞功能性单抗,为治疗肺癌提供靶向治疗剂,并为分离获得肺癌相关的分子靶标打下基础。方法从新鲜人肺癌组织分离肿瘤细胞免疫Balb/c小鼠,免疫小鼠脾细胞与SP2/0细胞融合后接种于甲基纤维素,制备大容量功能性抗体库。采用活细胞荧光、ELISA、免疫组化、肿瘤增殖、侵袭、黏附、动物体内治疗实验等方法筛选鉴定抑制肿瘤细胞的功能性单抗。结果本次融合共获得了1573株杂交瘤克隆,在能与肺癌细胞膜反应的314株克隆中,154株与正常肺组织不反应或低反应。功能性筛选发现41株单抗显著地抑制肺癌细胞增殖,24株能抑制肺癌细胞对Matrigel的侵袭,15株能抑制肺癌细胞与CollagenI的黏附,进一步实验验证了2株单抗能在体内抑制肺癌移植瘤的生长。结论采用大容量功能性抗体库技术成功获得了多株具有抑制肺癌细胞恶性生物学行为的功能性单抗,其中2株可能具有靶向治疗肺癌的应用潜力。

Objective Monoclonal antibodies which inhibit lung cancer may be candidate drugs for the antimalignancy therapy. Methods Malignant cells were separated form lung carcinoma tissues to immunize Balb/c mouse. The spleen cells of the immunized mouse were fused with SP2/I） cells and culture on the methyl cellulose. Antibodies of each clone were screened by immunofluorescence, immunohistochemistry, ELISA, cell proliferation assay, tumor cell invasion assay and adhesion assay. Functional antibodies were also studied by tumor treatment experiments in viva. Results One thousand five hundred and seventy-three monoclonal antibody clones were obtained by the fuse of spleen cells with SP2/0 cells. Fortyone clones could significantly suppress the lung cancer cell proliferation, 24 monoclonal antibody could inhibit tumor cell invasion in Matrigel, and 15 monoclonal antibody could decrease the adherence between lung cancer cells and Collagen I. In vivo assay further demonstrated that two of these functional antibodies could significantly inhibit lung tumor growth in mice. Conclusion By using functional antibody library, we successfully obtained functional monoclonal antibodies which suppressed the lung cancer cells in vitro and in vivo. These antibodies may be applied in anti-tumor therapy of lung carcinoma.