Monoclonal Antibodies to VP1 Recognize a Broad Range of Enteroviruses

• Published in: Journal of Clinical Microbiology 2009-10, Vol.47 (10), p.3108-3113
• Main Authors: Miao, Lynn Yihong, Pierce, Christina, Gray-Johnson, Jennifer, DeLotell, Jill, Shaw, Carl, Chapman, Nate, Yeh, Elaine, Schnurr, David, Huang, Yung T
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; Antibodies, Viral - immunology; Antibodies, Viral - isolation & purification; Applied microbiology; Biological and medical sciences; Capsid Proteins - genetics; Capsid Proteins - immunology; Cloning, Molecular; Conserved Sequence - immunology; Coxsackievirus B3; Cross Reactions; Enterovirus Infections - diagnosis; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Fundamental and applied biological sciences. Psychology; Gene Expression; Humans; Immunohistochemistry; Microbiology; Microscopy, Fluorescence; Poliovirus 1; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Sensitivity and Specificity; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects); Viral Structural Proteins - genetics; Viral Structural Proteins - immunology; Virology
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/jcm.00479-09

Enteroviruses (EVs) are common seasonal viruses that are associated with a variety of diseases. High-quality monoclonal antibodies (MAbs) are needed to improve the accuracy of EV diagnosis in clinical laboratories. In the present study, the full-length VP1 genes of poliovirus 1 (Polio 1) and coxsackievirus B3 (Cox B3) were cloned, and the encoded proteins were expressed and used as antigens in an attempt to raise broad-spectrum MAbs to EVs. Two pan-EV MAbs were isolated: one raised against Polio 1 VP1 and the other against Cox B3 VP1. The binding sites of both pan-EV MAbs were mapped to an amino acid sequence within a conserved region in the N terminus of Polio 1 VP1 by peptide and competition enzyme-linked immunosorbent assay. Two additional MAbs, an EV70-specific MAb and an EV71/Cox A16-bispecific MAb, developed against EV70 and 71 VP1 proteins, were pooled with the two pan-EV MAbs (pan-EV MAb mix) and tested for their sensitivity and specificity in the staining of various virus-infected cells. The pan-EV MAb mix detected all 40 prototype EVs tested and showed no cross-reactivity to 18 different non-EV human viruses. Compared with two commercially available EV tests, the pan-EV MAb mix exhibited higher specificity than one test and broader spectrum reactivity than the other. Thus, our study demonstrates that full-length Polio 1 VP1 and Cox B3 VP1 can serve as effective antigens for developing a pan-EV MAb and that the pan-EV MAb mix can be used for the laboratory diagnosis of a wide range of EV infections.