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用PCR技术从人外周血淋巴细胞中扩增出Fd段和κ链基因,构建了噬菌体抗体库,其库容量为1.5×10~8。将甲状旁腺素相关蛋白(PTHrP)合成肽固相化对该文库进行了“吸附-洗脱-扩增”的筛选,使特异性抗体富集了500多倍。经5轮筛选其噬菌体抗体阳性率为62%(31/50)。抑制实验表明。可溶性表达的Fab段具有结合PTHrP的特异性。SDS-PAGE分析显示其分子量约为50ku。
The Fd segment and kappa chain gene were amplified by PCR from human peripheral blood lymphocytes and the phage antibody library was constructed. The library capacity was 1.5 × 10 ~ 8. The library was “adsorbed-eluted-amplified” by immobilizing parathyroid hormone-related protein (PTHrP) synthetic peptide, enriching the specific antibody by more than 500-fold. After 5 rounds of screening, the phage antibody positive rate was 62% (31/50). Suppression experiments show. The solublely expressed Fab fragment has the specificity of binding to PTHrP. SDS-PAGE analysis showed that its molecular weight was about 50ku.