Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in Nicotiana benthamiana and Immunological Response in Mice

Author:

Jung Jae-Wan12ORCID,Park Pil-Gu3ORCID,Lee Won-Kyung1,Shin Jun-Hye12,Jang Mi-Hwa12,Seo Eun-Hye34ORCID,An Timothy3,Kim Young Beom5,Moon Myeong Hee5ORCID,Choi Seuk-Keun6,Yun Jee Sun6,Hong Kee-Jong3,Kim Seong-Ryong12

Affiliation:

1. Department of Life Science, Sogang University, Seoul 04107, Republic of Korea

2. PhytoMab Co., Seoul 04107, Republic of Korea

3. Department of Microbiology, Gachon University College of Medicine, Incheon 21936, Republic of Korea

4. BK21 Plus, Department of Cellular and Molecular Medicine, Konkuk University School of Medicine, Seoul 05029, Republic of Korea

5. Department of Chemistry, Yonsei University, Seoul 03722, Republic of Korea

6. Eubiologics Co., Seoul 06026, Republic of Korea

Abstract

The current production of the Japanese encephalitis virus (JEV) vaccine is based on animal cells, where various risk factors for human health should be resolved. This study used a transient expression system to express the chimeric protein composed of antigenic epitopes from the JEV envelope (E) protein in Nicotiana benthamiana. JEV multi-epitope peptide (MEP) sequences fused with FLAG-tag or 6× His-tag at the C- or N-terminus for the purification were introduced into plant expression vectors and used for transient expression. Among the constructs, vector pSK480, which expresses MEP fused with a FLAG-tag at the C-terminus, showed the highest level of expression and yield in purification. Optimization of transient expression procedures further improved the target protein yield. The purified MEP protein was applied to an ICR mouse and successfully induced an antibody against JEV, which demonstrates the potential of the plant-produced JEV MEP as an alternative vaccine candidate.

Funder

Korea Health Industry Development Institute

Ministry of Health & Welfare, Republic of Korea

Publisher

MDPI AG

Subject

Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis

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