Xia Jinsong, Bi Hao, Yao Qin, Qu Shen, Zong Yiqiang
Department of Nuclear Medicine, Tongji Hospital, Tongli Medical College, Huazhong Universty of Science and Technology, Wuhan 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2006;26(5):497-9. doi: 10.1007/s11596-006-0502-y.
In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E. coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 x 10(9) cfu/microg was obtained. After amplification with helper phage, the titer of antibody library reached 5 x 10(12) cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.
为构建抗卵巢肿瘤的单链抗体可变区（ScFv）噬菌体展示文库，采用RT-PCR从卵巢肿瘤患者淋巴细胞中扩增出人重链可变区基因（VH）和轻链可变区基因（VL），随后通过重叠延伸PCR（SOE）组装成ScFv基因。将所得ScFv基因电转化至大肠杆菌TG1中，并与辅助噬菌体M13KO7共感染进行扩增，以获得噬菌体展示文库。检测所得文库的库容和滴度。获得了库容约为3×10⁹ cfu/μg的噬菌体抗体文库。经辅助噬菌体扩增后，抗体文库滴度达到5×10¹² cfu/mL。成功构建了抗卵巢肿瘤的人ScFv文库，为筛选用于卵巢肿瘤放射免疫显像诊断的卵巢肿瘤特异性ScFv奠定了基础。
