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Clinical and Vaccine Immunology, August 2006, p. 953-957, Vol. 13, No. 8
1071-412X/06/$08.00+0     doi:10.1128/CVI.00037-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Human Neutralizing Fab Molecules against Severe Acute Respiratory Syndrome Coronavirus Generated by Phage Display

Xiaoping Kang, Bao-an Yang, Yuyang Hu, Hui Zhao, Wei Xiong, Yinhui Yang, Bingyin Si, and Qingyu Zhu*

State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China

Received 23 January 2006/ Returned for modification 16 March 2006/ Accepted 24 April 2006

Human recombinant Fab fragments specific for the spike protein of severe acute respiratory syndrome coronavirus (SARS-CoV) were screened from a human Fab library, which was generated from RNAs from peripheral lymphocytes of convalescent SARS patients. Among 50 randomly picked clones, 12 Fabs specially reacted with S protein by an enzyme-linked immunosorbent assay. The microneutralizing test showed that one clone, designated M1A, had neutralizing activity on Vero E6 cells against SARS-CoV. DNA sequence analysis indicated that the light- and heavy-chain genes of M1A Fab belong to the {kappa}2a and 4f families, respectively. A neutralizing test on purified M1A demonstrated that 0.5 mg/ml of M1A completely inhibited SARS-CoV activity, with an absence of cytopathic effect for 7 days. Real-time fluorescence reverse transcription-PCR also proved the neutralizing capacity of M1A. These data showed that the number of virus copies was significantly reduced in the M1A-treated group, suggesting an important role for M1A in passive immunoprophylaxis against the SARS virus.

* Corresponding author. Mailing address: Institute of Microbiology and Epidemiology, No. 20, Dong Da Jie, Fengtai District, Beijing 100071, China. Phone: 8610-63802193. Fax: 8610-63802193. E-mail: qingyzh{at}yahoo.com.cn.

Clinical and Vaccine Immunology, August 2006, p. 953-957, Vol. 13, No. 8
1071-412X/06/$08.00+0     doi:10.1128/CVI.00037-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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Antimicrob. Agents Chemother. Clin. Microbiol. Rev. Infect. Immun.
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Copyright © 2006 by the American Society for Microbiology. All rights reserved.