目的:筛选获取抗肺癌细胞膜的功能性单克隆抗体及其靶抗原，为肺癌靶向治疗提供候选治疗剂及靶标。方法: 将新鲜人肺癌组织细胞免疫BALB/c小鼠,采用脾细胞融合法制备大容量单克隆抗体库。建立肺癌细胞增殖、侵袭、迁移等定向功能差异细胞模型，采用活细胞免疫荧光与功能差异模型筛选其抗原在定向功能（细胞增殖、迁移、侵袭）差异模型细胞膜上差异表达的单抗，再经体外相应功能实验筛选具有抑制作用的功能性单抗，Western blotting鉴定这些单抗的抗原，裸鼠体内抑瘤实验鉴定部分单抗对肺癌的抑制作用。结果: 细胞融合后共获得2 893株杂交瘤克隆，其中与肺癌细胞膜结合反应的309株。分别建立了3种体外肺癌细胞增殖、侵袭、迁移差异模型用于筛选，筛选获得20株单抗能显著抑制肺癌细胞增殖、10株单抗能显著抑制肺癌细胞侵袭、5株单抗能显著抑制肺癌细胞迁移（P<0.05或P<0.01）。Western blotting鉴定了其中6株的抗原。选取其中2株单抗均能在裸鼠体内显著抑制肺癌移植瘤的生长（P<0.05或P<0.01）。结论: 采用大容量功能性单抗库技术结合功能差异细胞模型可批量筛选获得具有体内外抑制肺癌细胞恶性生物学行为的功能性单抗，为筛选肺癌靶向治疗剂及靶标提供了一种潜在的新方法。

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 functionaloriented 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 functionaloriented 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<001）. 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 functionaloriented 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 antimalignancy therapy.

国家重点基础研究发展计划（973）资助项目（No.2009CB521804）;国家高技术研究发展计划（863）资助项目（No.2006AA02Z479）。