INFLUENZA VACCINE Belgrade, Serbia March 27-29th, 2012 INSTITUTO / FUNDAÇÃO BUTANTAN SÃO PAULO - BRAZIL 1901: Vital Brazil creates Instituto Butantan and starts to produce anti-pest serum. Main producer of vaccines and sera in South America Production: ~ 160 million doses of vaccine (51 %) ~ 410,000 ampoules of serum (56 %) ( in 2010) Instituto Butantan | 1 INDUSTRIAL PRODUCTION SEASONAL INFLUENZA VACCINE 1999: Technology transfer signed with Butantan and Sanofi- Pasteur (Pasteur-Merieux) Quality control tests → Filled Vaccine → Formulated bulk → Blending, filling and packing → Production Technology 2009: Water Batches (11 lots) 2010: H3N2 production tests (59 lots) Seasonal vaccine 2010-11: ~ 5 millions doses (45 lots) 2011-12: ~ 20 millions doses (146 lots) Instituto Butantan | 2 INFLUENZA LABORATORY Instituto Butantan | 3 INFLUENZA PILOT PLANT Pilot Plant (renovated building): ~ 300 m2, BSL 3 Grantees: WHO, CDC, MoH Instituto Butantan | 4 PILOT SCALE PRODUCTION • 2007: testing production line - A/Panama/2007/99 (H3N2) • 2007- 08: Production - A/Vietnam/1194/2005- NIBRG-14 (H5N1): master and working seed and production (split and whole virus) • 2008: Installation of Quality Control Laboratory for Influenza • 2009-10: Production - A/California/210/2009 (A/H1N1): master and working seed and production (split and whole virus) • 2011: Production - A/Victoria/120/2009 (H3N2) and B/Brisbane/60/2008: master and working seed for seasonal vaccine Instituto Butantan | 5 INFLUENZA PILOT PLANT Instituto Butantan | 6 BUTANTAN INSTITUTE BUTANTAN FOUNDATION INFLUENZA VACCINE SPLIT X WHOLE VIRUS Instituto Butantan | 7 SPLIT X WHOLE STEP SPLIT WHOLE VIRUS Clarification X X X X Concentration X X X 1st purification X X X 2nd purification X Diafiltration/concentration X X Chromatograph* X X Spliting X Inactivation X X X X Final filtration X X X X * Size exclusion (sheparose 4 FF) Instituto Butantan | 8 SPLIT X WHOLE Fig.1. Flow diagram for the production of split and whole vaccine Alantoic fluid Clarification Concentration Clarification Purification Inactivation and diafiltration/concentration Split, diafiltration and inactivation Gel filtration chromatography Final filtration Final filtration SPLIT VACCINE WHOLE VACCINE C. Miyaki et al. / Vaccine 28 (2010) 2505–2509 Instituto Butantan | 9 WHOLE VIRUS VACCINE • Clarification: 7000 rpm/20 min or continuous flow • Inactivation: 0.2 % formaldehyde • Clarification: 7000 rpm/10 min or continuous flow • Concentration: 20 times • Ultrafiltration/Diafiltration: 20 times (PBS) • SEC: Sepharose 4FF High ~ 60 cm • Load 6 – 10 % CV • Final filtration: 0.65 + 0.45 + 0.22 µm Instituto Butantan | INFLUENZA WHOLE VIRUS VACCINE 97 66 45 30 20 14 AF W W S MW S W H3 H3 SDS-Page Instituto Butantan | 11 SPLIT X WHOLE H1N1 Split 1.5 20.8 41.0 2.0 H1N1 Whole 6.9 25.4 5.2 0.2 H3N2 Split 2.0 - - - H3N2 Whole 3.0 - - - B Split 1.3 - - - B Whole 1.5 - - - - On analysis Instituto Butantan | 12 SPLIT X WHOLE Instituto Butantan | 13 PHASE I CLINICAL TRIAL – A/H1N1 Instituto Butantan | Muito Obrigada! Cosue Miyaki Mauricio Meros de Oliveira [email protected] [email protected] Av. Vital Brasil, 1500 - Butantã São Paulo – SP Zip Code - 05503-900 (+ 55 11) 2627-9372 www.butantan.gov.br Instituto Butantan | 15 .