Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro

Zhong, Ying; Tang, Xiaoqian; Sheng, Xiuzhen; Xing, Jing; Zhan, Wenbin

doi:10.3390/ijms19092536

Open AccessArticle

Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro

by

Ying Zhong

¹,

Xiaoqian Tang

¹,

Xiuzhen Sheng

^1,*,

Jing Xing

¹ and

Wenbin Zhan

^1,2

¹

Laboratory of Pathology and Immunology of Aquatic Animals, KLMME, Ocean University of China, Qingdao 266003, China

²

Function Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China

^*

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2018, 19(9), 2536; https://doi.org/10.3390/ijms19092536

Submission received: 20 July 2018 / Revised: 22 August 2018 / Accepted: 25 August 2018 / Published: 27 August 2018

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

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Abstract

In previous research, a 32 kDa protein in lymphocystis disease virus (LCDV) was identified as viral attachment protein (VAP) that specifically interacted with the 27.8 kDa cellular receptor from flounder Paralichthys olivaceus gill (FG) cells, and the recombinant VAP (rVAP) was expressed in Escherichia coli strain BL21 (DE3). In this study, monoclonal antibodies (MAbs) against 32 kDa VAP are produced by immunization of BALB/c mice with the rVAP. Seven hybridoma secreting MAbs were screened by enzyme-linked immunosorbent assay, five of which designated as 1C6, 1C8, 3B5, 3D11 and 3H10 are cloned by the limiting dilution method, depending on the strongly positive results of ELISA. Western blotting analysis shows that the five MAbs can specifically react with the 32 kDa protein of LCDV and the purified 50 kDa rVAP, and the subtype of the MAbs is identified as IgG. Immunofluorescence results demonstrate that the specific fluorescence signals for LCDV appear in the cytoplasm of FG cells at 24 h post LCDV infection. Neutralization assay results indicate that pre-incubations of LCDV with the five MAbs can significantly decrease the LCDV copy numbers and delay the development of the cytopathic effect in FG cells, revealing that the five MAbs can neutralize the LCDV particles and block viral infection in vitro. The neutralizing MAbs against 32 kDa VAP would be useful for the study on the LCDV–host interaction and might be promising inhibitors of LCDV infection in fish.

Keywords: lymphocystis disease virus; monoclonal antibody; viral attachment protein; neutralization; flounder (Paralichthys olivaceus)

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MDPI and ACS Style

Zhong, Y.; Tang, X.; Sheng, X.; Xing, J.; Zhan, W. Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro. Int. J. Mol. Sci. 2018, 19, 2536. https://doi.org/10.3390/ijms19092536

AMA Style

Zhong Y, Tang X, Sheng X, Xing J, Zhan W. Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro. International Journal of Molecular Sciences. 2018; 19(9):2536. https://doi.org/10.3390/ijms19092536

Chicago/Turabian Style

Zhong, Ying, Xiaoqian Tang, Xiuzhen Sheng, Jing Xing, and Wenbin Zhan. 2018. "Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro" International Journal of Molecular Sciences 19, no. 9: 2536. https://doi.org/10.3390/ijms19092536

APA Style

Zhong, Y., Tang, X., Sheng, X., Xing, J., & Zhan, W. (2018). Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro. International Journal of Molecular Sciences, 19(9), 2536. https://doi.org/10.3390/ijms19092536

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Development and Characterization of Monoclonal Antibodies to the 32 kDa Viral Attachment Protein of Lymphocystis Disease Virus and Their Neutralizing Ability in Vitro

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