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

Original text

In some countries, a test for virus inactivation is carried out by inoculating 10 mice intracerebrally with 0.03ml of the final product.

New text

Original text

Each final lot shall be tested for abnormal toxicity by appropriate tests in mice and guinea pigs.

New text

Original text

A sample from each final lot shall be tested for potency. The challenge strain, reference material and test procedure shall be approved by the national control authority (see section B.1), which shall also set the minimum acdceptable potency.

New text

Development of appropriate in vitro methods validated for replacing animal models is strongly encouraged. If an in vitro assay has been developed, it should be implemented as the potency test if approved by the NRA. However, if an in vivo assay is carried out instead of an in vitro assay, it must be scientifically justified and should be approved by the NRA. If several final lots are issued from one final bulk product, the in vivo assay should be carried out on the final bulk product and omitted on the final lots in order to reduce animal use. After the demonstration of consistency of production by the in vivo assay on an appropriate number of final bulk products, a single dilution assay approved by the NRA should be carried out.

Original text

Each final lot shall be shown to be free from pyrogenic substances. The test shall be approved by the national control authority.

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 recombinant vaccine production strain (parental cell transformed with the recombinant expression construct) should be fully described and information should be given on the results of any adventitious agent testing required, and on the homogeneity and accuracy of the inserted sequence (including copy number per cell) for the MCB and WCB.

New text

Original text

At the end of the examination period, the cells should also be tested for haemadsorbing viruses

New text

At the end of the observation period a fraction of culture comprising not less than 25% of the total should be tested for the presence of haemadsorbing viruses, using red blood cells from guinea-pig or other suitable red blood cells. It is not necessary to use red blood cells from multiple species. If the red blood cells have been stored prior to use in the haemadsorption assay, the duration of storage should not have exceeded 7 days and the temperature of storage should have been in the range of 2–8 °C.

Original text

Each adsorbed antigen bulk should be tested for bacterial endotoxins using a method 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

The potency of each formulated final bulk before filling should be assessed by an appropriate in vivo method. If an in vivo potency test is used to test final fill lots, this test may be omitted on the formulated final bulk before filling. The methods used for antibody detection in the in vivo test and for the analysis of data should be approved by the NRA. The vaccine potency should be compared with that of a reference preparation approved by the NRA. In vitro methods such as ELISA may be developed to assess potency. With
the approval of the NRA, the in vitro assay may replace the in vivo assay when appropriately validated and when consistency of production is demonstrated.

New text

In vitro assays for antigen detection (e.g. ELISA) have been developed and are considered to be appropriate for the potency assay. A quantitative in vitro test, approved by the NRA and using appropriately characterized antibodies, should be performed using samples representative of each final vaccine bulk or final lot. If an in vivo assay is carried out instead of an in vitro assay, it must be scientifically justified and should be approved by the NRA.

Original text

A potency test should be carried out on each final lot as outlined above in section A.7.1.5. However, if the in vivo potency test has been performed on the final formulated bulk, the test on the final lot may be omitted, subject to the agreement of the NRA.

New text

A potency test should be carried out on each final lot as outlined above in section A.7.1.5. However, if a potency test has been performed on the final formulated bulk, the test on the final lot may be omitted, subject to the agreement of the NRA.

Original text

Each final lot should be tested for pyrogenic substances. Where appropriate, tests for endotoxin – for example, the limulus amoebocyte lysate (LAL) test – should be performed. However, where there is interference in the test (for example, from the adjuvant) a test for pyrogens in rabbits should be performed. A suitably validated monocyte-activation test may also be considered as an alternative to the rabbit pyrogen test. The test is conducted until consistency of production is demonstrated, subject to the agreement of 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.