[Construction and screening of human phage antibody library against SARS virus].
OBJECTIVE
To construct human phage antibody library against SARS virus.


METHODS
Human Fab genes were amplified by RT-PCR from lymphocytes of a convalescent SARS patient and cloned into vector pComb3 to construct phage antibody library. Antibodies against SARS virus were screened by biopanning with immobilized virus antigen. The binding specificity of phage antibody to SARS virus was detected by ELISA.


RESULTS
13 Ig genes were obtained by PCR. A human phage antibody library consisting of 1.3 x 10(6) clones was constructed by electrotransformation and 60 percent of the clones contained Fab genes. After three rounds of panning with SARS virus antigen, phage antibodies against SARS virus were specifically enriched. ELISA detection verified 10 positive phage antibodies that were highly specific for SARS virus.


CONCLUSIONS
Human phage antibody library against SARS virus has been constructed successfully, from which 10 anti-SARS virus antibodies were obtained.
Kang
Xiao-Ping
XP
The Army Center of Microbial Detection and Research, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China.
Xiong
Wei
W
Chang
Guo-Hui
GH
Zhu
Qing-Yu
QY
D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics* / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics*; D002417:Cattle; D006801:Humans; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D019151:Peptide Library