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Antiviral Res. 2017 Aug;144:153-163. doi: 10.1016/j.antiviral.2017.06.013. Epub 2017 Jun 19.

Construction of a hepatitis B virus neutralizing chimeric monoclonal antibody recognizing escape mutants of the viral surface antigen (HBsAg).

Author information

1
Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
2
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
3
Institute of Virology, Technical University of Munich/Helmholtz Zentrum München, Germany.
4
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
5
Department of Oncology-Pathology, Immune and Gene Therapy Lab, Cancer Center Karolinska (CCK), Karolinska University Hospital Solna and Karolinska Institute, Stockholm, Sweden; Department of Immunology, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran.
6
Section of Molecular Virology, Paul-Ehrlich-Institut, Langen, Germany.
7
Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: fshokri@tums.ac.ir.

Abstract

Hepatitis B virus (HBV) infection is a global burden on the health-care system and is considered as the tenth leading cause of death in the world. Over 248 million patients are currently suffering from chronic HBV infection worldwide and annual mortality rate of this infection is 686000. The "a" determinant is a hydrophilic region present in all antigenic subtypes of hepatitis B surface antigen (HBsAg), and antibodies against this region can neutralize the virus and are protective against all subtypes. We have recently generated a murine anti-HBs monoclonal antibody (4G4), which can neutralize HBV infection in HepaRG cells and recognize most of the escape mutant forms of HBsAg. Here, we describe the production and characterization of the chimeric human-murine antibody 4G4 (c-4G4). Variable region genes of heavy and light chains of the m-4G4 were cloned and fused to constant regions of human kappa and IgG1 by splice overlap extension (SOE) PCR. The chimeric antibody was expressed in Chinese Hamster Ovary (CHO)-K1 cells and purified from culture supernatant. Competition ELISA proved that both antibodies bind the same epitope within HBsAg. Antigen-binding studies using ELISA and Western blot showed that c-4G4 has retained the affinity and specificity of the parental murine antibody, and displayed a similar pattern of reactivity to 13 escape mutant forms of HBsAg. Both, the parental and c-4G4 showed a comparably high HBV neutralization capacity in cell culture even at the lowest concentration (0.6μg/ml). Due to the ability of c-4G4 to recognize most of the sub-genotypes and escape mutants of HBsAg, this antibody either alone or in combination with other anti-HBs antibodies could be considered as a potent alternative for Hepatitis B immune globulin (HBIG) as an HBV infection prophylactic or for passive immunotherapy against HBV infection.

KEYWORDS:

Chimeric anti-HBs monoclonal antibody; HBV neutralization; Viral escape

PMID:
28641998
DOI:
10.1016/j.antiviral.2017.06.013
[Indexed for MEDLINE]

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