摘要
目的构建抗人B细胞淋巴瘤单链抗体(ScFv)库。方法从人B细胞淋巴瘤细胞系Raji细胞免疫的BALB/c小鼠脾细胞中提取mRNA,RT-PCR扩增重、轻链可变区基因片断,连接成ScFv基因并扩增,将其克隆到噬菌体载体pHEN1中,构建成单链噬菌体抗体库。结果抗体库容量为3.4×106,约100%的噬菌体基因中有ScFv基因的插入。结论成功构建了抗人B细胞淋巴瘤噬菌体单链抗体库,方便进一步筛选抗人B细胞淋巴瘤抗体。
Objective To construct a single chain Fv （SeFv） phage display library against human B-lymphoma. Methods The mRNA was extracted from the spleen cells of the immunized BALB/c mice with Raji B-lymphoma cell strain and cDNA was synthesized by reverse transcriptase. The heavy-chain and light-chain variable region genes（VH and VL）of imm. unoglobulin were amplified by PCR and joined by a DNA linker to encode peptide as a single chain Fv（ScFv） fragment. ScFv frag ments were amplified and cloned into the phagemid vector pHEN1 and the phage display library was constructed, Results The ScFv phage display library had a capacity of approximately 3.4× 10^6 , with 100% of the phagemids containing ScFv gene insertion as demonstrated by PCR. Conclusion The ScFv phage display library against human B-lymphoma was successfully constructed. The present study can be used for succeeding screening of specific antibody against human B-lyrnphoma.
出处
《江苏医药》
CAS
CSCD
北大核心
2006年第1期40-42,T0002,共4页
Jiangsu Medical Journal
基金
国家自然科学基金(30400111)
江苏省自然科学基金(BK2004041)
关键词
B细胞淋巴瘤
单链抗体
噬菌体展示文库
B-lymphoma
Single chain Fv fragment（ScFv）
Phage display library
