INFLUENZA

Belgrade, Serbia March 27-29th, 2012 INSTITUTO / FUNDAÇÃO BUTANTAN -

1901: creates Instituto Butantan and starts to produce anti-pest serum. Main producer of 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 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 ) • 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 % • 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