2. 02/06/20151 COMBINED VACCINES WITH A HEP B COMPONENT The role of non-clinical testing in ensuring their safety & efficacy Roland Dobbelaer, Dr. Sc., Head Biological Standardisation Scientific Institute of Public Health Brussels Copyright, 1996 © Dale Carnegie & Associates, Inc.

3. 02/06/20152 COMBINED VACCINES ARE NOT NEW: DISEASE COMBOS Men A,C Men A,C,W,Y Pneu 23 Pneu conj 7, 11, 14 Flu A,A,B OPV 1, 2, 3

4. 02/06/20153 COMBINED VACCINES ARE NOT NEW: VIRAL COMBOS Me Mu Ru MMR MMR+V HepA/HepB

5. 02/06/20154 COMBINED VACCINES ARE NOT NEW: BACTERIAL + VIRAL COMBOS DT D-HepB DTP a,w / Hib DTP a,w - HepB / Hib DTP a,w -IPV 1,2,3 / Hib DTP a,w -IPV 1,2,3 -HepB/ Hib

6. 02/06/20155 PhEur VACCINES FOR HUMAN USE Vaccina ad usum humanum  “For a combined vaccine, where there is no monograph to cover a particular combination, the vaccine complies with the monograph for each individual component, with any necessary modifications approved by the competent authority.”

7. 02/06/20156 COMBINED VACCINES: Issues

8. 02/06/20157 Combining is more than mixing Formulation issues of combined vaccines  pH  Degree of adsorption  Ionic strength & Osmolality  Concentration and compatibility of:  adjuvant  buffer salts  antimicrobials  residual CHOH, A-biotics, endotoxins,...

9. 02/06/20158 PhEur Characterisation HBsAg –Complete protein, lipid and carbohydrate structure –Morphological characteristics of particles (electron microscopy) –Buoyant density (gradient centrifugation) –Antigenic epitopes –Primary structure (amino-acid composition & sequence analysis, peptide mapping)

10. 02/06/20159 PhEur HepB testing of culture and harvest and on purified antigen  CULTURE AND HARVEST –Identity, microbial purity, plasmid retention and consistency of yield  PURIFIED ANTIGEN –Total protein –Antigen content and identy (RIA, ELISA with MCl Ab against protective epitope) –Antigen/protein ratio

11. 02/06/201510 PhEur HepB testing of culture and harvest and on purified antigen  PURIFIED ANTIGEN (ctd.) –Molecular weight –Antigenic purity (>= 95 % HBsAg). –Composition (proteins, lipids, NA and CH) –Host-cell- and vector-derived DNA (<=10 pg/shd) –Caesium and/or other chemicals –Sterility

12. 02/06/201511 PhEur HepB testing of the final bulk and final lot  Final bulk –Antimicrobial preservative –Sterility  Final lot –Identity, Aluminium, Free formaldehyde, Antimicrobial preservative, Sterility, Pyrogens –Assay: either in vivo, by comparing its capacity to induce specific antibodies against hepatitis B surface antigen (HBsAg) in mice or guinea-pigs with the same capacity of a reference preparation, or in vitro, by an immunochemical determination of the antigen content.

13. 02/06/201512 PhEu rHepB in-vivo assay: mouse immunogenicity –Vaccinate –Incubate –Titrate –Calculate       >   

14. 02/06/201513 PhEur HepB in-vitro assay: antigen content (ELISA)  

15. 02/06/201514

16. 02/06/201515 TESTING TO ENSURE CONSISTENCY What if possible interactions between antigens are observed? –HepB –Di & Te without Pw –Te & Hib Te conj. –Hib & Pa  Compare with clinical reference and monitor consistency

17. 02/06/201516 IMPORTANCE OF  Research into correlates/surrogates for protection and defining significance of observed differences  Defining how much info has to be available before licensing and what can be left to post- licensing  Close epidemiological monitoring of vaccine coverage and efficacy & adverse reactions, pathogen circulation upon introduction of new (combined) vaccines  Increased public sensitivity to safety standards

18. 02/06/201517 Quality Surveillance of Biological Medicinal Products

19. 02/06/201518 BASIS FOR MUTUAL RECOGNITION QUALITY  THE WORLD ACCORDING TO EN45001  EDQM “Joint audits”  EDQM proficiency studies  WHO audits

20. 02/06/201519 CONTROL AUTHORITY BATCH RELEASE  PRODUCER-INDEPENDENT OMCL PRE-MARKETING –RE-testing and critical RE-evaluation of production & control protocol  to verify & monitor conformity with MAA, Ph. Eur. & WHO

21. 02/06/201520 CONTROL AUTHORITY BATCH RELEASE PROCEDURE

22. 02/06/201521 CONTROL AUTHORITY BATCH RELEASE Hepatitis B (rDNA) Vaccine  On the bulk purified antigen:  Identity and purity  On the final lot:  Identity and Assay (the assay serves as an identity test)  If an in vitro assay is used to determine the antigen content, it must be done on the final lot.  If an in vivo assay  is used it is required only whenever a new final bulk has been used.