Easily purified baculovirus/insect-system-expressed recombinant hepatitis B virus surface antigen fused to the N- or C-terminus of polyhedrin

• Published in: Archives of virology 2022-02, Vol.167 (2), p.345-354
• Main Authors: Silva, Leonardo A., Camargo, Brenda R., Araújo, Ana Carolina, Batista, Taylice Leonel, Ribeiro, Bergmann M., Ardisson-Araújo, Daniel M. P.
• Format: Article
• Language: English
• Subjects: Animals; Antigens; Antigens, Surface; Baculoviridae - genetics; Biomedical and Life Sciences; Biomedicine; C-Terminus; Cell fusion; DNA viruses; Expression vectors; Fusion protein; Hepatitis B; Hepatitis B surface antigen; Hepatitis B Surface Antigens - genetics; Hepatitis B virus - genetics; Humans; Infectious Diseases; Insecta; Medical Microbiology; Occlusion bodies; Original Article; Polyhedrin; Protein folding; Protein purification; Proteins; Scanning electron microscopy; Virions; Virology
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-021-05305-6

Baculoviruses are circular double-stranded DNA viruses that infect insects and are widely used as the baculoviral expression vectors (BEVs), which provide a eukaryotic milieu for heterologous expression. The most frequently used vector is based on Autographa californica multiple nucleopolyhedrovirus (AcMNPV). However, purification of recombinant proteins produced using BEVs is laborious, time-consuming, and often expensive. Numerous strategies have been explored to facilitate purification of heterologous proteins, such as fusion with occlusion body (OBs)-forming proteins like polyhedrin (Polh). Baculoviruses produce OBs in the late stages of infection to protect the virion in the cellular environment, and the main protein responsible for OB formation is Polh. In this study, we investigated the effect of fusing the gene that encodes the surface antigen (S-HBsAg) of hepatitis B virus (HBV) to either the N- or C-terminus of the AcMNPV Polh. The production of recombinant viruses and recombinant proteins was confirmed, and the ability to form chimeric S-HBsAg-containing OBs was accessed by light and scanning electron microscopy of infected cells. The fusion was found to affect the shape and size of the OBs when compared to wild-type OBs, with the N-terminal fusion producing less-amorphous OBs than the C-terminal construct. In addition, the N-terminal construct gave higher levels of expression than the C-terminal construct. Quantitative and qualitative immunoassays with human serum or plasma antibodies against HBsAg showed that the two forms of the antigen reacted differently. Although both reacted with the antibody, the N-terminal fusion protein reacted with more sensitivity (2.27-fold) and is therefore more suitable for quantitative assays than the C-terminal version. In summary, the BEVs represents a promising tool for the production of reagents for the diagnosis of HBV infection.