Monoclonal antibodies to SARS-associated coronavirus (SARS-CoV): identification of neutralizing and antibodies reactive to S, N, M and E viral proteins

Ralph A Tripp; Lia M Haynes; Deborah Moore; Barbara Anderson; Azaibi Tamin; Brian H Harcourt; Les P Jones; Mamadi Yilla; Gregory J Babcock; Thomas Greenough; Donna M Ambrosino; Rene Alvarez; Justin Callaway; Sheana Cavitt; Kurt Kamrud; Harold Alterson; Jonathan Smith; Jennifer L Harcourt; Congrong Miao; Raj Razdan; James A Comer; Pierre E Rollin; Thomas G Ksiazek; Anthony Sanchez; Paul A Rota; William J Bellini; Larry J Anderson

doi:10.1016/j.jviromet.2005.03.021

Monoclonal antibodies to SARS-associated coronavirus (SARS-CoV): identification of neutralizing and antibodies reactive to S, N, M and E viral proteins

J Virol Methods. 2005 Sep;128(1-2):21-8. doi: 10.1016/j.jviromet.2005.03.021.

Affiliation

¹ National Centers for Infectious Diseases, Division of Viral and Rickettsial Diseases, Respiratory and Enteric Virus Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd. NE, Mailstop G-09, Atlanta, GA 30333, USA.

Abstract

Monoclonal antibodies (Mabs) against the Urbani strain of the SARS-associated coronavirus (SARS-CoV) were developed and characterized for reactivity to SARS-CoV and SARS-CoV S, N, M, and E proteins using enzyme-linked immunoabsorbent (ELISA), radioimmunoprecipitation, immunofluorescence, Western Blot and microneutralization assays. Twenty-six mAbs were reactive to SARS-CoV by ELISA, and nine were chosen for detailed characterization. Five mAbs reacted against the S protein, two against the M protein, and one each against the N and E proteins. Two of five S protein mAbs neutralized SARS-CoV infection of Vero E6 cells and reacted to an epitope within amino acids 490-510 in the S protein. While two of the three non-neutralizing antibodies recognized at second epitope within amino acids 270-350. The mAbs characterized should prove useful for developing SARS-CoV diagnostic assays and for studying the biology of infection and pathogenesis of disease.

Publication types

Evaluation Study

MeSH terms

Animals
Antibodies, Monoclonal / biosynthesis
Antibodies, Monoclonal / immunology*
Antibodies, Viral / biosynthesis
Antibodies, Viral / immunology*
Antibody Specificity*
Cell Line
Chlorocebus aethiops
Coronavirus M Proteins
Coronavirus Nucleocapsid Proteins
Epitope Mapping
Humans
Membrane Glycoproteins / immunology
Neutralization Tests
Nucleocapsid Proteins / immunology
Severe Acute Respiratory Syndrome / virology
Severe acute respiratory syndrome-related coronavirus / immunology*
Spike Glycoprotein, Coronavirus
Vero Cells
Viral Envelope Proteins / immunology
Viral Matrix Proteins / immunology
Viral Structural Proteins / immunology*
Viroporin Proteins

Substances

Antibodies, Monoclonal
Antibodies, Viral
Coronavirus M Proteins
Coronavirus Nucleocapsid Proteins
E protein, SARS coronavirus
M protein, SARS-CoV
Membrane Glycoproteins
Nucleocapsid Proteins
Spike Glycoprotein, Coronavirus
Viral Envelope Proteins
Viral Matrix Proteins
Viral Structural Proteins
Viroporin Proteins
spike glycoprotein, SARS-CoV
spike protein, mouse hepatitis virus