PURIFICATION AND EVALUATION OF THE BIOACTIVITY OF THE RECOMBINANT P54 ANTIGEN OF THE AFRICAN SWINE FEVER VIRUS EXPRESSED FROM Nicotiana benthamina

Nguyễn Thị Trà; Nguyễn Thu Giang; Hồ Thị Thương; Trịnh Thái Vy; Hoàng Thị Thu Hằng; Vũ Duy Thái Sơn; Lê Thị Trà My; Nguyễn Thị Thu Hiền; Lê Thị Thanh; Phạm Bích Ngọc

doi:10.34238/tnu-jst.8404

PURIFICATION AND EVALUATION OF THE BIOACTIVITY OF THE RECOMBINANT P54 ANTIGEN OF THE AFRICAN SWINE FEVER VIRUS EXPRESSED FROM Nicotiana benthamina

About this article

Received: 24/07/23 Revised: 30/08/23 Published: 13/09/23

Authors

1. Nguyen Thi Tra , Institute of Biotechnology – VAST; Graduate University of Science and Technology - VAST
2. Nguyen Thu Giang, Institute of Biotechnology – VAST
3. Ho Thi Thuong, Institute of Biotechnology – VAST; Graduate University of Science and Technology - VAST
4. Trinh Thai Vy, Institute of Biotechnology – VAST; Graduate University of Science and Technology - VAST
5. Hoang Thi Thu Hang, Institute of Biotechnology – VAST; Graduate University of Science and Technology - VAST
6. Vu Duy Thai Son, Institute of Biotechnology – VAST
7. Le Thi Tra My, Institute of Biotechnology – VAST
8. Nguyen Thi Thu Hien, Institute of Biotechnology – VAST
9. Le Thi Thanh, Institute of Biotechnology – VAST
10. Pham Bich Ngoc, Institute of Biotechnology – VAST; Graduate University of Science and Technology - VAST

Abstract

African swine fever is a dangerous infectious disease caused by ASFV (African swine fever virus) in pigs, with mortality rates up to 100%. Currently, no officially licensed commercial vaccine against African swine fever is on the market. p54 is considered one of the main target proteins for ASFV vaccine development and ASFV diagnostic kit production. In this study, the DNA sequence encoding the ectodomain of p54 of the VNUAHY-ASFV/Vietnam2019 strain was optimized for expression in Nicotiana benthamiana, then artificially synthesized and inserted into the pRTRA vector containing the trimer and oligomer motifs. Next, the entire expression cassettes were inserted into the pCB301 vector and successfully transformed into Agrobacterium tumefaciens for transient expression in Nicotiana benthamiana. The expression of recombinant p54 protein in plants was determined by Western blot. The better expression motif was selected for purification by Immobilized affinity chromatography, tested for the immune response in mice, and evaluated for ASFV-specific IgG antibody titer by ELISA reaction. P54-pII-tp was selected to purify and induce an immune response in mice. ELISA results showed that recombinant p54-pII-tp stimulated ASFV-specific antibody production in laboratory mice. This result is a premise for research and development of subunit vaccines and diagnostic kits against ASFV infection in Nicotiana benthamiana.

Keywords

Transient expression; A. tumefaciens; N. benthamiana; ASFV strain in Vietnam; Recombinant p54 protein

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DOI: https://doi.org/10.34238/tnu-jst.8404

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