人源性抗胶质瘤抗体基因的筛选、表达及活性鉴定

To construct new specific human antibody gene library with phage display technology，developing a new way to increase the productivity of the antibody. Methods Anti-glioma antibody was tested and screened by ELISA and immunohistochemical staining to confirm the existence of the antibody in some glioma patients, providing a practical basis for the construction of antibody gene library. RT-PCR was employed to proliferate human antibody gene. Human glioma ScFv genes were recombined by Cre-loxp system to produce the recombined phage-display antibody library. Results Human glioma antibody was found from the peripheral blood samples in 5 volunteers of glioma patients. Anti-human glioma antibody gene library with a capacity of 2x107 was successfully constructed with quite a high quality of phage display. Three specific clones with positive reactivity were screened. Conclusion The study indicates that construction of human glioma antibody gene library by phage display technology may provide a new strategy for the gene treatment of human gliomas.

分离脑胶质瘤患者外周血IgG和IgM抗体，用免疫吸附实验和Western Blot检测抗脑胶质瘤抗体；采集阳性患者外周血淋巴细胞，以噬菌体表面呈现技术构建人源性抗体的基因文库；采用Western Blot筛选抗人脑胶质瘤单链抗体，获得抗人脑胶质瘤单链抗体基因，使其在大肠杆菌系统表达并坚定表达产物的活性。为脑胶质瘤的抗体导向治疗提供理论和实验依据。