Cloning and Expression of HBcAg Using Food Grade Vector pNZ8149 in Lactococcus Lactis for the Development of HBV Therapeutic Vaccine

DOI

10.2991/978-94-6463-112-8_76How to use a DOI?

Keywords

HBV; CHB; HBcAg; pNZ8149; usp45; Lactococcus lactis NZ3900; nisin

Abstract

Hepatitis B virus (HBV) infection remains a major cause of chronic liver diseases which may develop into cirrhosis and hepatocellular carcinoma (HCC). Although HBV prophylactic vaccine is available, currently there is no complete cure for chronic hepatitis B (CHB). One of the alternatives for CHB treatment is therapeutic vaccine. The HBV core antigen (HBcAg) becomes a suitable candidate for HBV therapeutic vaccine since it is known as a potent inducer of B-cell and T-cell responses. In this study, we report the development of HBV therapeutic vaccine by expressing HBcAg derived from HBV subgenotype B3, which is one of the predominant sub-genotypes in Indonesia. A food grade expression vector pNZ8149 which contains nisin-controlled gene as an inducible promoter, was used to express HBcAg in Lactococcus lactis NZ3900. Moreover, signal peptide usp45 was added in the N-terminal of the gene of interest to increase protein secretion efficiency. The native HBV core gene was synthesized and successfully cloned into plasmid pNZ8149 and transformed into Lactococcus lactis NZ3900 using lactose as a selection marker. The expression of HBcAg was induced using 10 ng/mL nisin and confirmed using dot blot hybridization analysis. Finally, the target protein which is 21 kDa in size was obtained by partial purification using size exclusion chromatography. In conclusion, HBcAg was successfully cloned and expressed using a food grade expression vector pNZ8149 in Lactococcus lactis NZ3900. These findings will pave the development of therapeutic vaccine for CHB treatment in the future.

Copyright

© 2023 The Author(s)

Open Access

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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TY  - CONF
AU  - Rifqiyah N. Umami
AU  - Rahma I. Anwar
AU  - Hidayah Murtiyaningsih
AU  - Apon Z. Mustopa
PY  - 2023
DA  - 2023/03/01
TI  - Cloning and Expression of HBcAg Using Food Grade Vector pNZ8149 in Lactococcus Lactis for the Development of HBV Therapeutic Vaccine
BT  - Proceedings of the 1st International Conference for Health Research – BRIN (ICHR 2022)
PB  - Atlantis Press
SP  - 833
EP  - 846
SN  - 2468-5739
UR  - https://doi.org/10.2991/978-94-6463-112-8_76
DO  - 10.2991/978-94-6463-112-8_76
ID  - Umami2023
ER  -