WHO/IVB/11.11 Manual for Quality Control of Diphtheria, Tetanus and Pertussis Vaccines Immunization, Vaccines and Biologicals WHO/IVB/11.11 Manual for Quality Control of Diphtheria, Tetanus and Pertussis Vaccines Immunization, Vaccines and Biologicals The Department of Immunization, Vaccines and Biologicals thanks the donors whose unspecified financial support has made the production of this document possible. This document was published by the Expanded Programme on Immunization (EPI) of the Department of Immunization, Vaccines and Biologicals Ordering code: WHO/IVB/11.11 Printed: April 2013 This publication is available on the Internet at: www.who.int/vaccines-documents/ Copies of this document as well as additional materials on immunization, vaccines and biologicals may be requested from: World Health Organization Department of Immunization, Vaccines and Biologicals CH-1211 Geneva 27, Switzerland • Fax: + 41 22 791 4227 • Email: [email protected] • © World Health Organization 2013 All rights reserved. Publications of the World Health Organization can be obtained from WHO Press, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel: +41 22 791 3264; fax: +41 22 791 4857; email: [email protected]). 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Printed by the WHO Document Production Services, Geneva, Switzerland ii Contents Preface .............................................................................................................................. vii Chapter I: General ................................................................................................3 I.1 General introduction ........................................................................................3 I.1.1 Scope ....................................................................................................................3 I.1.2 Quality assurance ...............................................................................................3 I.1.3 References ............................................................................................................6 I.2. Standardization in vaccine testing .................................................................7 I.2.1 Introduction.........................................................................................................7 I.2.2 Quality control charts .........................................................................................7 I.3. Role of Biostatistics in Vaccine Potency Testing ..........................................9 I.4. The use of laboratory animals in the quality control of vaccines ............10 I.4.1 Environment .....................................................................................................10 I.4.2 Nutrition ............................................................................................................11 I.4.3 Choice of animals ..............................................................................................11 I.4.4 Microbiological quality .....................................................................................12 I.4.5 References ..........................................................................................................14 I.4.6 Selected further reading ....................................................................................14 I.5. General laboratory procedures .....................................................................15 I.5.1 The laboratory environment ............................................................................15 I.6. Laboratory safety ............................................................................................15 I.6.1 Protection for staff.............................................................................................16 I.6.2 Protection of the environment .........................................................................17 I.6.3 Selected further reading ....................................................................................18 iii Chapter II. Testing for Diphtheria Vaccines ..................................................21 II.1 Potency ..............................................................................................................21 II.1.1 General introduction ........................................................................................21 II.1.2 Potency in guinea pigs by challenge ................................................................24 II.1.3 Potency in guinea pigs by serology ...................................................................30 II.1.4 Potency in mouse by Vero cell assay ................................................................52 II.1.5 In vivo toxin neutralisation test .......................................................................55 II.2 Specific toxicity ................................................................................................60 II.2.1 In vivo test for absence of toxin and reversion to toxicity in guinea pigs .....60 II.2.2 Vero cell test for absence of toxin and reversion to toxicity ...........................63 II.3 Lf and identity .................................................................................................73 II.3.1 Flocculation test (Ramon and laser light scattering).......................................73 II.3.2 Radial Immunodiffusion ..................................................................................81 II.3.3 Rocket immunoelectrophoresis .........................................................................86 II.4 Antigen content and degree of adsorption..................................................91 II.4.1 Capture and direct ELISA ...............................................................................91 Chapter III: Testing for Tetanus vaccines .....................................................103 III.1 Potency: ..........................................................................................................103 III.1.1 General introduction ......................................................................................103 III.1.2 Potency in guinea pigs and mice by challenge (lethal and paralysis) ..........106 III.1.3 Potency in guinea pigs by serology .................................................................112 III.1.4 Potency in mice by serology (ToBI) ...............................................................130 III.1.5 In vivo toxin neutralisation test in mice ........................................................139 III.2 Specific toxicity ..............................................................................................144 III.2.1 In vivo test for absence of toxin and reversion to toxicity in guinea pigs ...144 III.3 Lf and identity ...............................................................................................147 III.3.1 Flocculation test (Ramon and laser light scattering).....................................147 III.3.2 Radial immunodiffusion .................................................................................155 III.3.3 Rocket immunoelectrophoresis .......................................................................160 III.4 Antigen content and degree of adsorption................................................165 III.4.1 Capture ELISA ...............................................................................................165 iv Chapter IV: Testing for whole cell pertussis vaccine ...................................175 IV.1 Kendrick test ..................................................................................................175 IV.2 Specific toxicity tests .....................................................................................187 IV.2.1 Mouse weight gain test (MWGT) ..................................................................187 IV.2.2 Other tests for monitoring pertussis toxicity .................................................192 IV.3 Methods used for strain characterization and identity tests .................219 IV.3.1 Slide agglutination