Monoclonal antibodies for diagnosis of enterovirus 71

Li Xu, Kao Jean Huang, Tzong-Shiann Ho, Chia Chyi Liu, Ying Ray Lee, Ching Yen Lin, David Shiuan, Xing Hong Jiang

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Enterovirus 71 (EV71), one of the major causative agents of hand, foot, and mouth disease (HFMD), is now recognized as an emerging neurotropic virus in Asia and may cause severe neurologic complications and mortalities. Laboratory diagnosis of EV71 infection must be efficient and accurate, which could be accomplished by various immunoassays. In this study we use a live EV71 isolate, Tainan/4643/98, with genotype C2 as an immunogen to sensitize BALB/c (H-2(d)) mice and then generate the EV71-specific murine monoclonal antibodies. Five hybridoma clones were established and their monoclonal antibodies were characterized. All five clones are applicable in immunofluorescence staining but with different sensitivities - that is, MAbs 22, 24, and 27 were sensitive in IFA detection, and MAbs 22 and 24 were also confirmed in flow cytometry. None of these cross-reacted with coxsackievirus A16 (CVA16) or Echovirus type 6 (ECHO6), but each varied in binding to different EV71 subgenogroups (B1, B4, B5, C2, and C4). Western blot analysis revealed that all of these MAbs reacted with EV71 VP1 capsid proteins, and in addition MAbs 22 and 24 exhibited potent neutralizing activities against EV71 and protected cells from infection. Further, mapping the epitopes for each MAb revealed that only MAb 27 showed positive for the linear epitope DVIESSIGDSVSRAL, which was located at the N-terminus (a.a. 6-20) of EV71 VP1 and highly conserved among all EV71 subgenotypes. Thus, these MAbs may provide valuable tools for the laboratory diagnosis of EV71 infection and for vaccine development.

Original languageEnglish
Pages (from-to)386-394
Number of pages9
JournalMonoclonal Antibodies in Immunodiagnosis and Immunotherapy
Volume32
Issue number6
DOIs
Publication statusPublished - 2013 Dec 1

Fingerprint

Enterovirus
Monoclonal Antibodies
Enterovirus Infections
Clinical Laboratory Techniques
Human echovirus 6
Clone Cells
Hand, Foot and Mouth Disease
Epitope Mapping
Hybridomas
Capsid Proteins
Immunoassay
Nervous System
Fluorescent Antibody Technique
Epitopes
Flow Cytometry
Vaccines
Western Blotting
Genotype
Staining and Labeling
Viruses

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Xu, Li ; Huang, Kao Jean ; Ho, Tzong-Shiann ; Liu, Chia Chyi ; Lee, Ying Ray ; Lin, Ching Yen ; Shiuan, David ; Jiang, Xing Hong. / Monoclonal antibodies for diagnosis of enterovirus 71. In: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. 2013 ; Vol. 32, No. 6. pp. 386-394.
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abstract = "Enterovirus 71 (EV71), one of the major causative agents of hand, foot, and mouth disease (HFMD), is now recognized as an emerging neurotropic virus in Asia and may cause severe neurologic complications and mortalities. Laboratory diagnosis of EV71 infection must be efficient and accurate, which could be accomplished by various immunoassays. In this study we use a live EV71 isolate, Tainan/4643/98, with genotype C2 as an immunogen to sensitize BALB/c (H-2(d)) mice and then generate the EV71-specific murine monoclonal antibodies. Five hybridoma clones were established and their monoclonal antibodies were characterized. All five clones are applicable in immunofluorescence staining but with different sensitivities - that is, MAbs 22, 24, and 27 were sensitive in IFA detection, and MAbs 22 and 24 were also confirmed in flow cytometry. None of these cross-reacted with coxsackievirus A16 (CVA16) or Echovirus type 6 (ECHO6), but each varied in binding to different EV71 subgenogroups (B1, B4, B5, C2, and C4). Western blot analysis revealed that all of these MAbs reacted with EV71 VP1 capsid proteins, and in addition MAbs 22 and 24 exhibited potent neutralizing activities against EV71 and protected cells from infection. Further, mapping the epitopes for each MAb revealed that only MAb 27 showed positive for the linear epitope DVIESSIGDSVSRAL, which was located at the N-terminus (a.a. 6-20) of EV71 VP1 and highly conserved among all EV71 subgenotypes. Thus, these MAbs may provide valuable tools for the laboratory diagnosis of EV71 infection and for vaccine development.",
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Xu, L, Huang, KJ, Ho, T-S, Liu, CC, Lee, YR, Lin, CY, Shiuan, D & Jiang, XH 2013, 'Monoclonal antibodies for diagnosis of enterovirus 71', Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, vol. 32, no. 6, pp. 386-394. https://doi.org/10.1089/mab.2013.0033

Monoclonal antibodies for diagnosis of enterovirus 71. / Xu, Li; Huang, Kao Jean; Ho, Tzong-Shiann; Liu, Chia Chyi; Lee, Ying Ray; Lin, Ching Yen; Shiuan, David; Jiang, Xing Hong.

In: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, Vol. 32, No. 6, 01.12.2013, p. 386-394.

Research output: Contribution to journalArticle

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AU - Xu, Li

AU - Huang, Kao Jean

AU - Ho, Tzong-Shiann

AU - Liu, Chia Chyi

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AU - Shiuan, David

AU - Jiang, Xing Hong

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