Monoclonal antibodies to VP1 recognize a broad range of enteroviruses.
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.
Miao
Lynn Yihong
LY
R&D Department, Diagnostic Hybrids, Inc., Athens, OH 45701, USA. miao@dhiusa.com
Pierce
Christina
C
Gray-Johnson
Jennifer
J
DeLotell
Jill
J
Shaw
Carl
C
Chapman
Nate
N
Yeh
Elaine
E
Schnurr
David
D
Huang
Yung T
YT
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000276:immunology; D003429:Cross Reactions; D004769:Enterovirus Infections / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins; C442540:VP1 protein, Poliovirus; D015678:Viral Structural Proteins