title: Development and preclinical testing of HNVAC, a cell culture-based H1N1 pandemic influenza vaccine from India.
abstract: Several limitations of the use of embryonated eggs and the threat of pandemics have highlighted the need for other platforms for the production of influenza vaccines. We report the indigenous development and pre-clinical testing of an MDCK-based H1N1 pandemic influenza vaccine HNVAC from India. The cell bank and virus seed were characterized extensively. The cells were characterized by PCR, electron microscopy, and karyotyping, and found to be of female canine epithelial origin. The virus was confirmed by neutralization, haemagglutination inhibition, neuraminidase inhibition, and PCR and nucleotide sequencing. Adventitious agent testing was performed by both in vitro and in vivo studies. The in vitro studies included culturing, haemadsorption, haemagglutination, PCR and RT-PCR, whereas in vivo studies included passage in embryonated eggs and in laboratory animals. Both cell bank and virus seed were free of adventitious agents. MDCK cell lysates as well as cellular DNA did not produce tumours in newborn or adult laboratory animals. The bioprocess parameters were standardized to recover antigen with minimal levels of process-related impurities. The vaccine bulk was tested for the presence of specific antigen, and quantified by single radial immunodiffusion. Finally, non-adjuvanted and aluminium hydroxide adjuvanted vaccine formulations were found to be safe in preclinical toxicity studies in mice, rats, guinea pigs and rabbits, and immunogenic in mice and rabbits. This is the first and only cell culture-based influenza vaccine platform developed in any developing country.
authors.affiliation: Ella Foundation, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India. Electronic address: hegden@ellafoundation.org.
authors.forename: Nagendra R
authors.initials: NR
authors.lastname: Hegde
authors.affiliation: Ella Foundation, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Ella Foundation, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: P Panduranga
authors.initials: PP
authors.lastname: Rao
authors.affiliation: Bharat Biotech International Limited, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: P Krishna
authors.initials: PK
authors.lastname: Kumari
authors.affiliation: Bharat Biotech International Limited, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: Yashpal
authors.initials: Y
authors.lastname: Kaushik
authors.affiliation: RCC Laboratories India Private Limited, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: R
authors.initials: R
authors.lastname: Ravikrishnan
authors.affiliation: RCC Laboratories India Private Limited, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: Sai D
authors.initials: SD
authors.lastname: Prasad
authors.affiliation: Bharat Biotech International Limited, Genome Valley, Turkapally, Shameerpet Mandal, Hyderabad 500078, India.
authors.forename: Krishna M
authors.initials: KM
authors.lastname: Ella
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004285:Dogs; D005260:Female; D006168:Guinea Pigs; D006385:Hemagglutination Inhibition Tests; D007194:India; D053118:Influenza A Virus, H1N1 Subtype; D007252:Influenza Vaccines / Q000096:biosynthesis* / Q000276:immunology*; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits; D051381:Rats; D015164:Vaccines, Inactivated / Q000096:biosynthesis / Q000276:immunology; D014776:Virus Cultivation
chemical_list: D007252:Influenza Vaccines; D015164:Vaccines, Inactivated
keywords: Cell culture; H1N1; Immunogenicity; Influenza; Pandemic; Pre-clinical; Safety; Vaccine