Construction of human ScFv phage display library against ovarian tumor.
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.
Xia
Jinsong
J
Department of Nuclear Medicine, Tongji Hospital, Tongli Medical College, Huazhong Universty of Science and Technology, Wuhan 430030, China.
Bi
Hao
H
Yao
Qin
Q
Qu
Shen
S
Zong
Yiqiang
Y
D061505:Cell Surface Display Techniques; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000235:genetics*
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies