349 results
| WHO guideline title | Product | TRS | Test name | Test category | 3Rs approach | Toggle to view all updates |
|---|---|---|---|---|---|---|
| Recommendations to assure the quality, safety and efficacy of DT-based combined vaccines |
DT-Combined vaccines
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980 Annex 6
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Reversion to toxicity of pertussis toxin
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Toxicity
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Original textSpecify the dates of the beginning and end of incubation, and the number, strain and sex of animals used – this test is not necessary for a product obtained by genetic modification New textShould be removed/revised based on part A text.
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| Recommendations to assure the quality, safety and efficacy of DT-based combined vaccines |
DT-Combined vaccines
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980 Annex 6
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If performed at the final lot stage: in vivo assay for D, T, aP and, depending on the marketing authorization, for IPV and HepB components
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Potency
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Original textSpecify the strain, sex, weight range and number of animals used; the dates, volumes, route and doses used for immunization and challenge or bleeding; the nature, lot number and potency of the reference vaccine; and the responses at each dose. New textRemove or revise based on new language for potency testing.
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| Recommendations to assure the quality, safety and efficacy of DT-based combined vaccines |
DT-Combined vaccines
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980 Annex 6
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Reversion to toxicity
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Toxicity
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Original textSpecify the dates of the beginning and end of incubation, the dates of the beginning and end of the test, the number of animals used, the volume inoculated into cell culture (for diphtheria only) or injected into animals, the number of animals used (if relevant), and the test results New textShould be removed/revised based on part A text.
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
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979 Annex 4
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Residual levels of endotoxin:
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Pyrogenicity/endotooxin testing
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MAT
rFC
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Original textthe antigens should be tested for residual endotoxin content by means of the Limulus amoebocyte lysate (LAL) test or other appropriate assay New textThe need for pyrogenicity testing should be assessed during the manufacturing development process and be re-evaluated following any significant changes in the production process or relevant reported production inconsistencies that may influence pyrogenicity. A risk-based approach should be implemented which is suitable to the manufacturing process and the product depending on the potential presence of endotoxins and non-endotoxin pyrogens. The endotoxin content of the final product should be determined using a suitable in vitro assay, such as the recombinant factor C (rFC) or limulus/tachypleus amoebocyte lysate (LAL/TAL) tests. The rFC method is strongly recommended due to concerns over the impact on the sustainability of limulus stocks. The endotoxin content should be consistent with levels found to be acceptable in final product lots used in clinical trials and within the limits agreed upon with the NRA. A monocyte activation test (MAT) may be used for pyrogen testing after a product-specific validation. The use of the rabbit pyrogen test should be avoided due to its inherent variability, high retesting rates, and interspecies differences in pyrogenic responses as compared to humans.ReferencesMAT:Schindler, S., von Aulock, S., Daneshian, M. and Hartung, T. (2009) “Development, validation and applications of the monocyte activation test for pyrogens based on human whole blood”, ALTEX - Alternatives to animal experimentation, 26(4), pp. 265–277. doi: 10.14573/altex.2009.4.265.
rFC: Biotechniques 2021 May;70(5):290-300. doi: 10.2144/btn-2020-0165. Epub 2021 May 6.
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
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979 Annex 4
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Residual activity of pertussis toxin
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Toxicity
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CHO cell assay
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Original textThe amount of residual biologically active PT in the individually or co-purified antigens should be estimated after detoxification by means of a sufficiently sensitive test such as the HIST or the CHO cell assay New textThe PTx inactivation process should be controlled and demonstrated to consistently reduce the active PTx to levels which are found to be safe in clinical trials. Inactivation of PTx can be monitored using a CHO cell clustering assay or similar in vitro method. The CHO cell clustering response has been demonstrated to have greater sensitivity and lower variability than the mouse histamine sensitization test (HIST) which is no longer recommended. Although a modified CHO cell clustering method can be used to monitor residual PTx activity in the presence of an adjuvant, inactivation of PTx should be controlled and verified prior to adsorption. The PTx inactivation process should be demonstrated during process development to produce a stable toxoid which does not undergo reversion during downstream processing steps or under recommended storage conditions. Subsequent testing for reversion of PTx should not be necessary once stability of the PTd has been demonstrated. The testing for residual PTx activity or activity from reversion is not necessary for PTd derived from genetic inactivation. The genetic insert should be confirmed to be stable, and the cell line shown to be absent of an active PTx gene.Referenceskataoka M et al. Chinese hamster ovary (CHO) cell clustering does not correlate with in vivo histamine-sensitization when measuring residual activity of aldehyde-treated pertussis toxin (PT). Biologicals, 2002, 30(4):297–302.
Ph Eur 2.6.33
Isbrucker R, Daas A, Wagner L, Costanzo A.Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines. Pharmeuropa Bio&SN | June 2016
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
|
979 Annex 4
|
Residual levels of endotoxin
|
Pyrogenicity/endotooxin testing
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MAT
rFC
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Original textResidual levels of endotoxin: the bulk material or antigens should be tested for residual endotoxin content by means of the LAL test or other appropriate assay. New textThe need for pyrogenicity testing should be assessed during the manufacturing development process and be re-evaluated following any significant changes in the production process or relevant reported production inconsistencies that may influence pyrogenicity. A risk-based approach should be implemented which is suitable to the manufacturing process and the product depending on the potential presence of endotoxins and non-endotoxin pyrogens. The endotoxin content of the final product should be determined using a suitable in vitro assay, such as the recombinant factor C (rFC) or limulus/tachypleus amoebocyte lysate (LAL/TAL) tests. The rFC method is strongly recommended due to concerns over the impact on the sustainability of limulus stocks. The endotoxin content should be consistent with levels found to be acceptable in final product lots used in clinical trials and within the limits agreed upon with the NRA. A monocyte activation test (MAT) may be used for pyrogen testing after a product-specific validation. The use of the rabbit pyrogen test should be avoided due to its inherent variability, high retesting rates, and interspecies differences in pyrogenic responses as compared to humans.ReferencesMAT:Schindler, S., von Aulock, S., Daneshian, M. and Hartung, T. (2009) “Development, validation and applications of the monocyte activation test for pyrogens based on human whole blood”, ALTEX - Alternatives to animal experimentation, 26(4), pp. 265–277. doi: 10.14573/altex.2009.4.265.
rFC: Biotechniques 2021 May;70(5):290-300. doi: 10.2144/btn-2020-0165. Epub 2021 May 6.
|
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
|
979 Annex 4
|
Residual activity of pertussis toxin
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Toxicity
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CHO cell assay
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Original textEach final bulk of vaccine should be tested for active PT using a HIST or another test that is sufficiently sensitive to detect the level of toxin activity agreed with the NRA New textThis test may not be needed at this stage if the test has been applied at an earlier stage. Otherwise use same text as A.3.3.
Referenceskataoka M et al. Chinese hamster ovary (CHO) cell clustering does not correlate with in vivo histamine-sensitization when measuring residual activity of aldehyde-treated pertussis toxin (PT). Biologicals, 2002, 30(4):297–302.
Ph Eur 2.6.33
Isbrucker R, Daas A, Wagner L, Costanzo A.Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines. Pharmeuropa Bio&SN | June 2016
|
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
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979 Annex 4
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Reversion to toxicity
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Toxicity
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CHO cell assay
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Original textAccelerated reversion testing, consisting of HIST performance on final bulk or the final lot incubated for at least four weeks at 37 °C, may be used to demonstrate that it is unlikely that the chemically inactivated PT will regain some of its toxicity before the vaccine expiry date. Some NRAs may not require this test for the release of each new lot but only as part of process validation. New textThis test may not be needed at this stage if the test has been applied at an earlier stage. Otherwise use same text as A.3.3.
Referenceskataoka M et al. Chinese hamster ovary (CHO) cell clustering does not correlate with in vivo histamine-sensitization when measuring residual activity of aldehyde-treated pertussis toxin (PT). Biologicals, 2002, 30(4):297–302.
Ph Eur 2.6.33
Isbrucker R, Daas A, Wagner L, Costanzo A.Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines. Pharmeuropa Bio&SN | June 2016
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
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979 Annex 4
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Immunological activity
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Potency
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MIA (Multiplex Immunological Assay)
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Original textTwo methods are currently used in the lot release procedure to assess the immunogenicity or potency in mice after vaccination with acellular pertussis vaccine: the mouse immunogenicity test (MIT) and the MICA. The development of alternative assays to MIT and MICA is encouraged. An alternative assay – the guinea-pig immunogenicity test – has been adopted in some countries New textAs biological assays (e.g. humoral antibody response in sera from a suitable species) with the titration of Ab by in vitro methods (in vitro TNT, ELISA, MIT depending on the component tested) and/or physicochemical tests have been developed and are considered to be more precise and reproducible than the challenge test, a biological and/or physicochemical assay should be used if approved by the NRA. In some countries the titration of Abs are performed using multiplex immunological methods for combined DTaP vaccines. If several final lots are issued from one final bulk product, the potency assay should be carried out on the final bulk product and omitted on the final lots. After the demonstration of consistency of production by the biological and/or physicochemical assay on an appropriate number of final bulk products, a single dilution assay approved by the NRA should be carried out. Biological and/or physicochemical assays are preferred. However, if an in vivo challenge test for Diphtheria, Pertussis & Tetanus is carried out, it must be scientifically justified and should be approved by the NRA. A single dilution assay may be implemented for Diphtheria, Pertussis & Tetanus components after demonstration of consistency of production on an appropriate number of final bulk products and should be approved by the NRA. In the context of a 3Rs strategy, development of a package of appropriately validated in vitro assays for the characterization of the drug product is strongly encouraged in order to replace animal models. For all components, in vitro antigenicity assays are being developed and may be considered as potency assays once they are appropriately validated.ReferencesDevelopment and validation of a robust multiplex serological assay to quantify antibodies specific to pertussis antigens Gowrisankar Rajam* , George Carlone1, Ellie Kim, Jin Choi, Simon Paulos, SoHee Park, Amilia Jeyachandran, Yamini Gorantla, Emily Wong, Amit Sabnis, Peter Browning, Rita Desai, Conrad P. Quinn, Jarad Schiffer
Biologicals. 2019 January ; 57: 9–20. doi:10.1016/j.biologicals.2018.11.001.
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| Recommendations to assure the quality, safety and efficacy of acellular pertussis vaccines |
Pertussis acellular vaccines
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979 Annex 4
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Innocuity test
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GST
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Original textEach final lot should be tested for unexpected toxicity (sometimes called abnormal toxicity or innocuity) by a method approved by the NRA. This test may be omitted for routine lot release once consistency of production has been well established to the satisfaction of the NRA New textThis needs to be removed as per WHO TRS 1016, 2019 page No 32-33
ReferencesBiologicals. 2020 Jul; 66: 17–20.
doi: 10.1016/j.biologicals.2020.05.003
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