DIGANTA BARMAN, RAMAR KRISHNAMURTHY, R. RAVISHANKARAN AND MAITRI SHUKLA
C. G. Bhakta Institute of Biotechnology, Uka Tarsadia University, Maliba Campus, Mahuva-Bardoli Rd, Tarsadi, Bardoli, Tarsadi-394 620 (Gujarat), India
*(e-mail: Digantabarman99@gmail.com; Mobile: 87238 66259)
(Received: January 15, 2025; Accepted: February 20, 2025)
ABSTRACT
Two hundred fifty million people suffer from chronic liver disease due to the hepatitis B virus (HBV), leading to cirrhosis and hepatocellular cancer. Despite a preventive vaccine, coverage is still not sufficient and antiviral medications are often ineffective. Monoclonal antibodies (mAbs) are crucial for a targeted therapeutic approach, enhancing the ability to control and eliminate the virus. These antibodies can neutralize the virus, prevent its spread and assist the immune system in combating the illness, particularly for persistent HBV patients. In this study, peripheral blood mononuclear cells (PBMC) from 10 hepatitis B vaccinated healthy donors, lymphoblastoid cell lines (LCLs) were generated following transformation with the Epstein-Barr virus (EBV). Subsequently, clones secreting antibodies to hepatitis B surface antigen (anti-HBs antibodies) were screened. Following this, 9 LCL clones and three clones fused with mouse myeloma were generated, and the resulting human anti-HBs monoclonal antibody was purified from the supernatant. These purified antibodies were biotinylated, and the most suitable pairs for sandwich ELISA were identified. The efficacy of these antibody pairs was tested using clinical samples positive for the Hepatitis B virus. The test demonstrated a detection sensitivity of 100% and a specificity of 99.3%. Thus, diagnostic possibility using hetero-hybridoma mAbs was verified and further study can be carried out to affirm its therapeutic potential.
Key words: Hepatitis B virus, lymphoblastoid cell lines, peripheral blood mononuclear cells, sandwich ELISA