Review of animal use requirements in WHO biologics guidelines – database of suggested guideline revisions

Original text

Each final lot should be tested in mice or guinea-pigs to confirm the absence of abnormal toxicity using a test approved by the NRA. This test may not be required for routine lot release after the consistency of production has been established to the satisfaction of the NRA.

New text

Original text

The vaccine in the final container should be tested for pyrogenic activity through intravenous injection into rabbits. A Limulus amoebocyte lysate (LAL) test may be used in lieu of the rabbit pyrogen test if it has been validated. Similarly, a suitably validated monocyte activation test may be considered as an alternative to the pyrogen test.
When testing is performed on RTS,S reconstituted with the adjuvant system, the rabbit test for pyrogens should be performed because the adjuvant may interfere with the LAL test.

New text

The 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.

Original text

The characteristics of the recombinant production strain (host cell in combination with the expression vector system) should be fully described, and information should be given about the absence of adventitious agents (35, 36) and gene homogeneity for the MCBs and WCBs. ....The MCB and WCB should be tested for the absence of adventitious agents according to Part A of WHO General requirements for the sterility of biological substances no. 6 (1973) (35) or by a method approved by the NRA.

New text

Original text

Each final purified antigen bulk should be tested for bacterial endotoxins. The method and the concentration limits used should be approved by the NRA.

New text

The 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.

Original text

In vivo assay.
Number of mice tested:
Species, strain, sex, ages and weight range:
Dates of immunization:
Date of test:
Vaccine doses (dilutions) and number of animals responding at each dose:

New text

Original text

A suitable in vitro or in vivo assay for immunogenicity or antigenicity should therefore be concidered.

New text

In vitro assays, such as monoclonal antibody ELISAs, are likely to be suitable for the routine testing of synthetic peptide vaccines.

Original text

The product specification should also include tests to ensure that the final dosage form complies with the usual safety tests, such as pyrogenicity and sterility, appropriate to a parenteral preparation, together with tests of identity, antigen content and general innocuity.

New text

The 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.

Original text

The pyrogen content of the final bulk shall be determined by a method agreed with the NCA.

New text

The 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.

Original text

The innocuity of the final product shall be tested parenterally in guinea-pigs and mice by a method approved by the national control authority.

New text

Original text

Each final lot shall be tested for pyrogenic substanes by a method approved by the NCA

New text

The 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.