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Published on: January 17, 2015
[Construction of phage antibody library for Fab fragment from a convalescent patient infected with SARS coronavirus]
Jie Wu1, Wen-ling Zheng, Hong-bin Zhang
1Institute of Cancer Molecular Biology,Guangzhou Command General Hospital, Guangzhou 510010, China. wujie55@hotmail.com
Aim:
To construct the phage antibody library for Fab fragment from a convalescent patient infected with SARS coronavirus.
Methods:
Total RNA was extracted from peripheral blood lymphocytes of a patient with SARS virus-specific IgG antibody and was transcribed reversely into cDNA. Using the cDNA as template, Fd and light chain of Fab genes were amplified by PCR with specific primer and were cloned into phagemid pComb3. Then the recombinant phagemid was electroporated into E.coli XL-1 Blue. The recombinant rate was detected by enzyme digestion analysis, and library repertoire was determined.
Results:
The antibody library of Fab fragment from a convalescent patient infected with SARS conronavirus was constructed. The recombinant rate of light chain and heavy chain of Fd gene were 91% and 75%, respectively. The library size was 7.23 x 10(7).
Conclusion:
Phage antibody library for human Fab fragment has been constructed successfully, which lays the foundation for further study.
