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The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation

Enterovirus 71 (EV71) is the major etiological agent contributing to the development of hand-foot-mouth disease (HFMD). There are not any global available vaccines or antibody drugs against EV71 released yet. In this study, we perform the virus immunization in a cost-effective and convenient approac...

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Autores principales: Lee, Chi-Hsin, Huang, Peng-Nien, Mwale, Pharaoh Fellow, Wang, Wei-Chu, Leu, Sy-Jye, Tseng, Sung-Nien, Shih, Shin-Ru, Chiang, Liao-Chun, Mao, Yan-Chiao, Tsai, Bor-Yu, Dlamini, Nhlanhla Benedict, Nguyen, Tien-Cuong, Tsai, Chen-Hsin, Yang, Yi-Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9653933/
https://www.ncbi.nlm.nih.gov/pubmed/36361757
http://dx.doi.org/10.3390/ijms232112967
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author Lee, Chi-Hsin
Huang, Peng-Nien
Mwale, Pharaoh Fellow
Wang, Wei-Chu
Leu, Sy-Jye
Tseng, Sung-Nien
Shih, Shin-Ru
Chiang, Liao-Chun
Mao, Yan-Chiao
Tsai, Bor-Yu
Dlamini, Nhlanhla Benedict
Nguyen, Tien-Cuong
Tsai, Chen-Hsin
Yang, Yi-Yuan
author_facet Lee, Chi-Hsin
Huang, Peng-Nien
Mwale, Pharaoh Fellow
Wang, Wei-Chu
Leu, Sy-Jye
Tseng, Sung-Nien
Shih, Shin-Ru
Chiang, Liao-Chun
Mao, Yan-Chiao
Tsai, Bor-Yu
Dlamini, Nhlanhla Benedict
Nguyen, Tien-Cuong
Tsai, Chen-Hsin
Yang, Yi-Yuan
author_sort Lee, Chi-Hsin
collection PubMed
description Enterovirus 71 (EV71) is the major etiological agent contributing to the development of hand-foot-mouth disease (HFMD). There are not any global available vaccines or antibody drugs against EV71 released yet. In this study, we perform the virus immunization in a cost-effective and convenient approach by preparing virus particles from size exclusion and immunization of chicken. Polyclonal yolk-immunoglobulin (IgY) was simply purified from egg yolk and monoclonal single-chain variable fragments (scFv) were selected via phage display technology with two scFv libraries containing 6.0 × 10(6) and 1.3 × 10(7) transformants. Specific clones were enriched after 5 rounds of bio-panning and four identical genes were classified after the sequence analysis. Moreover, the higher mutation rates were revealed in the CDR regions, especially in the CDR3. IgY showed specific binding activities to both EV71-infected and Coxsackievirus 16-infected cell lysates and high infectivity inhibitory activity of EV71. However, while IgY detected a 37 kDa protein, the selected scFv seemingly detected higher size proteins which could be cell protein instead of EV71 proteins. Despite the highly effective chicken antibody generation, the purity of virus particles prepared by size exclusion is the limitation of this study, and further characterization should be carried out rigorously.
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spelling pubmed-96539332022-11-15 The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation Lee, Chi-Hsin Huang, Peng-Nien Mwale, Pharaoh Fellow Wang, Wei-Chu Leu, Sy-Jye Tseng, Sung-Nien Shih, Shin-Ru Chiang, Liao-Chun Mao, Yan-Chiao Tsai, Bor-Yu Dlamini, Nhlanhla Benedict Nguyen, Tien-Cuong Tsai, Chen-Hsin Yang, Yi-Yuan Int J Mol Sci Article Enterovirus 71 (EV71) is the major etiological agent contributing to the development of hand-foot-mouth disease (HFMD). There are not any global available vaccines or antibody drugs against EV71 released yet. In this study, we perform the virus immunization in a cost-effective and convenient approach by preparing virus particles from size exclusion and immunization of chicken. Polyclonal yolk-immunoglobulin (IgY) was simply purified from egg yolk and monoclonal single-chain variable fragments (scFv) were selected via phage display technology with two scFv libraries containing 6.0 × 10(6) and 1.3 × 10(7) transformants. Specific clones were enriched after 5 rounds of bio-panning and four identical genes were classified after the sequence analysis. Moreover, the higher mutation rates were revealed in the CDR regions, especially in the CDR3. IgY showed specific binding activities to both EV71-infected and Coxsackievirus 16-infected cell lysates and high infectivity inhibitory activity of EV71. However, while IgY detected a 37 kDa protein, the selected scFv seemingly detected higher size proteins which could be cell protein instead of EV71 proteins. Despite the highly effective chicken antibody generation, the purity of virus particles prepared by size exclusion is the limitation of this study, and further characterization should be carried out rigorously. MDPI 2022-10-26 /pmc/articles/PMC9653933/ /pubmed/36361757 http://dx.doi.org/10.3390/ijms232112967 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lee, Chi-Hsin
Huang, Peng-Nien
Mwale, Pharaoh Fellow
Wang, Wei-Chu
Leu, Sy-Jye
Tseng, Sung-Nien
Shih, Shin-Ru
Chiang, Liao-Chun
Mao, Yan-Chiao
Tsai, Bor-Yu
Dlamini, Nhlanhla Benedict
Nguyen, Tien-Cuong
Tsai, Chen-Hsin
Yang, Yi-Yuan
The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title_full The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title_fullStr The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title_full_unstemmed The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title_short The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation
title_sort bottlenecks of preparing virus particles by size exclusion for antibody generation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9653933/
https://www.ncbi.nlm.nih.gov/pubmed/36361757
http://dx.doi.org/10.3390/ijms232112967
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