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SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity.
Biomed Environ Sci. 2005 Dec; 18(6):363-74.BE

Abstract

OBJECTIVE

To develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection.

METHODS

By using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined.

RESULTS

After four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus.

CONCLUSION

The SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection.

Authors+Show Affiliations

State Key Laboratory for Infectious Disease Control and Prevention, Institute for Viral Disease Control and Prevention, Beijing 100052, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16544518

Citation

Liang, Mi-Fang, et al. "SARS Patients-derived Human Recombinant Antibodies to S and M Proteins Efficiently Neutralize SARS-coronavirus Infectivity." Biomedical and Environmental Sciences : BES, vol. 18, no. 6, 2005, pp. 363-74.
Liang MF, Du RL, Liu JZ, et al. SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity. Biomed Environ Sci. 2005;18(6):363-74.
Liang, M. F., Du, R. L., Liu, J. Z., Li, C., Zhang, Q. F., Han, L. L., Yu, J. S., Duan, S. M., Wang, X. F., Wu, K. X., Xiong, Z. H., Jin, Q., & Li, D. X. (2005). SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity. Biomedical and Environmental Sciences : BES, 18(6), 363-74.
Liang MF, et al. SARS Patients-derived Human Recombinant Antibodies to S and M Proteins Efficiently Neutralize SARS-coronavirus Infectivity. Biomed Environ Sci. 2005;18(6):363-74. PubMed PMID: 16544518.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity. AU - Liang,Mi-Fang, AU - Du,Run-Lei, AU - Liu,Jing-Zhi, AU - Li,Chuan, AU - Zhang,Quan-Fu, AU - Han,Lu-Lu, AU - Yu,Jian-Shi, AU - Duan,Shu-Min, AU - Wang,Xiao-Fang, AU - Wu,Kong-Xing, AU - Xiong,Zhao-Hui, AU - Jin,Qi, AU - Li,De-Xin, PY - 2006/3/21/pubmed PY - 2006/5/18/medline PY - 2006/3/21/entrez SP - 363 EP - 74 JF - Biomedical and environmental sciences : BES JO - Biomed Environ Sci VL - 18 IS - 6 N2 - OBJECTIVE: To develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection. METHODS: By using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined. RESULTS: After four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus. CONCLUSION: The SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection. SN - 0895-3988 UR - https://www.unboundmedicine.com/medline/citation/16544518/SARS_patients_derived_human_recombinant_antibodies_to_S_and_M_proteins_efficiently_neutralize_SARS_coronavirus_infectivity_ L2 - http://www.diseaseinfosearch.org/result/6415 DB - PRIME DP - Unbound Medicine ER -