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

Chi-Hsin Lee, Peng-Nien Huang, Pharaoh Fellow Mwale, Wei-Chu Wang, Sy-Jye Leu, Sung-Nien Tseng, Shin-Ru Shih, Liao-Chun Chiang, Yan-Chiao Mao, Bor-Yu Tsai, Nhlanhla Benedict Dlamini, Tien-Cuong Nguyen, Chen-Hsin Tsai, Yi-Yuan Yang

研究成果: 雜誌貢獻文章同行評審

摘要

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 × 106 and 1.3 × 107 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.
原文英語
期刊International journal of molecular sciences
23
發行號21
DOIs
出版狀態已發佈 - 10月 2022

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