Isolation of specific humanized anti-D-dimer scFv fragments from scFv phage libraries

VernacularTitle:从scFv噬菌体库分离特异性的人源化抗D-dimer抗体
Author: Hongli XIA ; Zui TAN ; Dejie CHEN ; Jianguo QIAO ; Renfeng QIU
Publication Type:Journal Article
Keywords: Screening; Anti-D-dimer antibody; Phage display
From: Chinese Journal of Microbiology and Immunology 2011;31(2):168-172
CountryChina
Language:Chinese
Abstract: Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.