Serum Institute of India Pvt. Ltd. Live attenuated Influenza vaccine Parikshit Tyagi, MSc, PhD Sr. Manager, Quality Control
Introduction founded in 1966 by Dr. Cyrus Poonawalla with the aim of manufacturing life-saving immuno-biologicals, which were in shortage in the country and imported at high prices. facilities to meet the internationally accepted environmental standards are state-of-the-art and comply with BSL-2 and BSL-2+ standards. one of the largest suppliers of vaccines to over a 140 countries and it is estimated that about 65% of the children in the world receive at least one vaccine manufactured by Serum Institute of India.
Products commercial & pipeline BCG & Onco BCG vaccine DPT group vaccine Bacterial vaccines Haemophilus type b Conjugate MMR group Vaccines Viral Vaccines Influenza vaccine (Pandemic & seasonal) Rabies vaccine Poliomyelitis Vaccine Conjugate vaccine Meningococcal A Conjugate Vaccine (Lyophilised) Recombinant & combination vaccines Penta vac- DTP (WC),Hepatitis-B (rdna) & Haemophilus Type b Conjugate Vaccine (Adsorbed) Q Vac-DTP (WC), Haemophilus Type b Conjugate Vaccine (Adsorbed) Diphtheria, Tetanus, Pertussis & Hepatitis-B Vaccine (Adsorbed) Hepatitis-B Vaccine (rdna) Product in pipeline Rota virus vaccine Dengue vaccine HPV, DTaP Pentavalent meningococcal conjugate vaccine Pneumococcal conjugate vaccine RMAB/DMAB
Influenza Vaccine
Foundation of Project Considering the complete absence of seasonal influenza vaccine uptake and a large scale investment required to setup the influenza vaccine manufacturing unit, SIIPL started its activity to address the need for pandemic influenza vaccine with active collaboration and support from World Health Organization.
Choice of Vaccines Reported influenza vaccine technologies Attenuated influenza vaccine for immunization through nasal route Inactivated vaccine whole virus/subunit virus preparations Large number of doses in a short duration Small manufacturing setup Low cost Widely used methodology
Pandemic Influenza vaccine H1N1 (2009) Inactivated influenza vaccine Live attenuated influenza vaccine Limited number of doses Large number of doses Pregnant women and children below 3 years of age All individuals above 3 years of age
Challenges overcome Facility: Separate facility for IIV and LAIV, larger setup-egg incubators, TFF systems, Ultra-centrifuge. Formulation: IIV- Whole virus/subunit, adjuvantation LAIV-Liquid/Freeze dried, stabilizer, spray device. Licensing: Demonstration of safety for IIV and Immunogenicity of LAIV.
Manufacturing of LAIV Working seed Inoculation of 9-11 days old SPF eggs Harvesting of egg allantoic fluid concentration by diafiltration Formulation of batch with predetermined set titre Final bulk Filling/Lyophilization/ labeling Addition of stabilizer Clarified Monovalent virus pool Drug substance Final vaccine Drug product
Live attenuated A/H1N1 influenza vaccine challenge studies in ferrets Ferret Immunized with LAIV through intranasal route Challenged intratracheally with 10 6 TCID 50 of wildtype H1N1 on day 28 post immunization Euthanasia- Day 4 post challenge Decreased body weight loss, Low relative lung weight, Absence of fever, Little or no (<5%) affected lung parenchyma Viral load decreased or absent in vaccinated animals as compared to control animals Lung pathology showed reduced severity of H1N1 in vaccinated animals High HAI and MN antibody titres in vaccinated animals as compared to control animals
Live attenuated A/H1N1 influenza vaccine challenge studies in ferrets Placebo-treated control group Severely affected, Dark hyperaemic & consolidated lung Vaccinated group No evident discernable lesions. Normal aerated lung tissue is bright pink in color
Phase-I Study Double-blind, randomized, placebo-controlled study 50 healthy adults of 18-49 years of age Safety findings Most of the reactions were mild. All reactions resolved within 3 days without any sequalae. No solicited adverse event/sae. No change in hematology, biochemical, electrolyte and urine parameters. No case of Guillian-Barre syndrome, Bell s palsy or any other chronic condition
Phase-II/III Study Double-blind, randomized, placebo-controlled 5 Study Centers across India Four assays used: Haemagglutination inhibition Microneutralization Serum IgG Mucosal IgA
Comparison between Russian & SIIPL LAIV
License for LAIV was received on 3rd July 2010 and for IIV on 6th August 2010. More than 2.5 million doses of LAIV distributed all over India.
Initial LAIV packaging
Seasonal LAIV (egg based) Commercial tri LAIV approved by DCG(I) and WHO PQ Cloning & characterization of new master donor virus (MDV) as per WHO/EU/CBER guidelines Safety in vitro & in vivo, respiratory pathogens, whole genome sequencing etc QLAIV liquid vaccine established formulation & shelf life,
Challenges Ferret challenge study for quadrivalent formulation Bridging study to switch from tri to quadrivalent liquid formulation Exploring qpcr/ddpcr based potency assay where specific homologues antisera is not required for QLAIV
Cell culture based LAIV Characterization of MDCK cell bank Deep sequencing for extraneous agents Whole genome sequencing Tumorigenicity & TPD 50 (subcutaneous/intranasal) Oncogenicity with ras/myc plasmids as positive control In vivo & in vitro extraneous agents MAP, PERT/ TEM
Challenges Comparative efficacy of egg and cell culture based QLAIV in ferrets Clinical trails Formulation Shelf life estimation Dossier submission to various regulatory authorities
Special thanks Dr Marie-Paule Kieny and WHO team Dr Larisa Rudenko and IEM team Dr John Wood and NIBSC team Dr A.C. Mishra, Ex-Director, NIV, Pune Dr Surendra Singh and staff at DCG(I) and Central Drug Lab., India PATH, BARDA team USFDA
Colleagues Dr. Suresh Jadhav, Executive Direction, RA & QA Dr. Rajeev Dhere, Executive Director, Vaccines Dr. Sunil Gairola, Director, QC Dr. Leena Yeolekar, Consultant Mr Vivek Vaidya, Additional Director, Production Mr S.G. Bankar, Additional Director, QC Mr. Milan Ganguly, Manager, Production
Thanks