Synthesis of Human Antibodies Against HBsAg in Newly Established Chinese Hamster Lung (CHL-YN) Cell Line
DOI:
https://doi.org/10.5614/j.math.fund.sci.2023.54.3.1Keywords:
antibodies, hepatitis B, CHL-YN cells, transient expression, vector plasmidAbstract
Hepatitis B immunoglobulin (HBIG) is an effective treatment for hepatitis B, including postexposure prophylaxis of HBV infection, prevention of HBV reinfection in liver transplant patients, and reducing sexual transmission. This study investigated the synthesis of human IgG antibodies that specifically target HBsAg subtype adr in CHL-YN cells, a newly established cell line that grows faster than CHO-K1. To achieve the synthesis of human IgG antibodies, a plasmid vector encoding DNA sequences for human IgG antibodies against HBsAg was constructed and then transiently transfected into CHL-YN cells. The expression and antigen-binding capacity of the recombinant human IgG antibodies were analyzed using western blot and ELISA. The results showed successful expression and secretion of human IgG antibodies that recognize HBsAg subtype adr in CHL-YN cells. The ELISA test confirmed the specificity of the human IgG antibodies towards HBsAg subtype adr. Thus, this study concluded that human IgG antibodies that target HBsAg subtype adr were transiently expressed in CHL-YN cells.
References
World Health Organization. Hepatitis B. https://www.who.int/news-room/fact-sheets/detail/hepatitis-b. 2020.
Departemen Kesehatan Indonesia. Situasi Dan Analisis Hepatitis. http://www.depkes.go.id/resources/download/pusdatin/infodatin/infodatin-hepatitis.pdf. 2016
Centers for Disease Control and Prevention. Recommendation of the Immunization Advisory Committee (ACIP) Postexposure prophylaxis of Hepatitis B. MMWR, 21, pp. 285-90, 1984.
U. S Food and Drug Administration. Hepatitis B Immunoglobulin. 2021. https://www.ftc.gov/system/files/documents/cases/181_0081_c4654_grifols-biotest_analysis.pdf.
Stricker, E.A., Tiebout, R.F., Lelie, P.N., Zeijlemaker, W.P., A human monoclonal IgG1 lambda anti- hepatitis B surface antibody. Production, properties, and applications. Scand J Immunol., 22 pp. 337-343, 1985.
Desgranges, C., Paire, J., Pichoud, C., Souche, S., Frommel D. & Trepo C., High affinity human monoclonal antibodies directed against hepatitis B surface antigen. J Virol Methods, 16, pp. 281- 292, 1987.
Sa?adu, A., Loeniskar, M., Bidwel, D., Howard, C., McAdam, K.P. & Voller, A., Development and characterization of human anti-HBs antibodies. J Virol Methods, 37, pp. 1-11. 1992.
Matsui K., Safety and neutralizing activity against HBV of a human monoclonal anti- HBS antibody. J Nihon Univ Med Assoc, 12, pp. 1163- 1171. 1982
Maeda, F., Nagatsuka, Y., Ihara, S., Aotsuka, S., Ono, Y., Inoko, H., Takekoshi, M., Bacterial expression of human recombinant monoclonal antibody fab fragment against hepatitis B surface antigen., J Med Virol., 4, pp. 338-345 1984.
Hempel, F., Lau, J., Klingl, A. & Maier, U.G., Algae as protein factories: expression of a human antibody ant the respective antigen in the diatom Phaeodactylum tricronutum. PLoS One, 12, pp. e28424. 2011.
Shanmugaraj, B. Kaewta R., Suwimon, M., Arunee, T. & Waranyoo, P., Monoclonal Antibodies B38 and H4 Produced in Nicotiana benthamiana SARS-CoV-2 in vitro. Front Plant Sci., 11, pp. 589995. 2020.
Vanier, G., Hempel, F., Chan, P., Rodamer, M., Vaudry, D., Maier & U.G., Lerouge, P. et al., Biochemical characterization of human anti- hepatitis B monoclonal antibody produced in microalgae phaedactylum tricornutum. PLoS ONE, 10, e0139282. 2015.
Yamano-Adachi, N., Arishima, R., Puriwat, S. & Omasa, T., Establishment of fast-growing serum-free immortalizes cells from Chinese hamster lung tissue for biopharmaceutical production. Sci Rep., (1), pp. 17612. 2020.
Li, Wenzhou, Kerwin, J.L., Schiel, J.E., Mouchachoir, C.A., Davis, D., Mahan, A. & Benchaar, S.A., Structural elucidation of post- translational modification in monoclonal antibodies. ACS Symposium Series, 1201, pp. 119-183. 2015.
Liu, H. & May, K., Disulfide bond structures of IgG molecules: structural variations, chemical modifications and possible impacts to stability and biological function. MAbs, 1, pp. 17-23. 2012.
De Marco, A., Recombinant antibody production evolves into multiple options aimed at yielding reagents suitable for application- specific needs. Microbial Cell Factories 1, pp. 125, 2015.
Komatsu, K., Kumon, K., Arita, M., Onitsuka, M., Omas, T. & Yohda, M., Effect of the disulfide isomerase PDIa4 on the antibody production of Chinese hamster ovary cells, J Biosci Bioeng, 6, pp. 637- 643, 2020.
