Construction of a hepatitis B virus neutralizing chimeric monoclonal antibody recognizing escape mutants of the viral surface antigen (HBsAg)
Autor: | Mohammad Mehdi Amiri, Motahareh Bahadori, Samira Farid, Sebastian Maximilian Altstetter, Mohammad Hojjat-Farsangi, Jalal Khoshnoodi, Ulrike Protzer, Fazel Shokri, Mahmood Jeddi-Tehrani, Michael Chudy, Tohid Kazemi, Forough Golsaz-Shirazi, Lisa S. Wolff, Felix Bohne |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Hepatitis B virus HBsAg medicine.drug_class Recombinant Fusion Proteins Blotting Western Enzyme-Linked Immunosorbent Assay CHO Cells Antibodies Viral Monoclonal antibody medicine.disease_cause Virus Epitope Epitopes Mice 03 medical and health sciences Cricetulus 0302 clinical medicine Antigen Virology medicine Animals Humans Pharmacology Hepatitis B Surface Antigens Hepatitis B immune globulin biology Antibodies Monoclonal virus diseases Antibodies Neutralizing digestive system diseases 030104 developmental biology biology.protein Mutant Proteins 030211 gastroenterology & hepatology Antibody Protein Binding medicine.drug |
Zdroj: | Antiviral Research. 144:153-163 |
ISSN: | 0166-3542 |
Popis: | 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. |
Databáze: | OpenAIRE |
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