[Construction of phage antibody library for Fab fragment from a convalescent patient infected with SARS coronavirus].
OBJECTIVE
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).


CONCLUSIONS
Phage antibody library for human Fab fragment has been constructed successfully, which lays the foundation for further study.
Wu
Jie
J
Institute of Cancer Molecular Biology,Guangzhou Command General Hospital, Guangzhou 510010, China. wujie55@hotmail.com
Zheng
Wen-ling
WL
Zhang
Hong-bin
HB
Wang
Jie
J
Jiang
Yue-hua
YH
Huang
Wen-jie
WJ
Ma
Wen-li
WL
D000914:Antibodies, Viral / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D050438:Genes, Immunoglobulin Heavy Chain; D050439:Genes, Immunoglobulin Light Chain; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007146:Immunoglobulin lambda-Chains / Q000235:genetics; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000276:immunology*
D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D019151:Peptide Library