1. Clinical Laboratory, Affiliated Foshan Maternity & Child Healthcare Hospital, Southern Medical University, Foshan, 528000, China;
2. School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China;
3. Guangzhou Bioneeds Biotechnology CO., LTD, Guangzhou, 510000, China;
4. Department of Nuclear Medicine, Yuxi People's Hospital of Yunnan Province,Yuxi, 653100, China;
5. Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, 21205, USA;
6. South China Institute of Biomedicine, Guangzhou, 510000, China.
# These authors contributed equally to this work.
Rabies continues to be a huge threat to public health. The rabies virus envelope glycoprotein (RABV G) is a major rabies virus antigen and contains neutralizing epitopes, which are primary candidates for subunit vaccines and diagnostic antigens. However, the production and purification of rRABV G while retaining its antigenic and immunogenic remains to be a challenge. Here, we aimed to establish a platform for rRABV G production and purification, and determine the immunogenicity and antigenicity of rRABV G. The cDNA fragment encoding the soluble form of RABV G was synthesized and cloned into a lentiviral expressing vector. Recombinant lentiviral vector LV-CMV-RABV G-eGFP was packaged, titered, and then transduced into HEK 293T cells. The cell culture supernatant was purified using nickel affinity chromatography and subsequently confirmed through Western Blot analysis and indirect enzyme-linked immunosorbent assay (ELISA). The ELISA utilized human sera obtained from individuals who had been vaccinated with the human commercial Purified Vero Cells Rabies Vaccine (PVRV). Notably, we observed a neutralizing antibody response in immunized pigs rather than in mice. This discrepancy could potentially be attributed to factors such as the instability of the rRABV G protein, variations in host responses, and variances in the adjuvant used. Taking all these findings into account, the rRABV G protein generated in this study exhibits promise as a potential vaccine candidate for the prevention of rabies.
Keywords: Rabies virus, Envelope glycoprotein, Secretory expression, Lentiviral vector, HEK 293T