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

Original text

Additional 10ml samples of each pool shall be tested in both human and simian cells.

New text

A strategy for testing adventitious viruses in vaccines must be developed based on a risk assessment. Relevant culture methods and/or specific molecular biology or broad molecular methods should be part of the overall testing package with the agreement of the NRA. In vivo tests may only be used if the risk assessment indicates that this test provides an additional risk mitigation taking into account the overall testing package.

Original text

Additional 10ml samples of each pool shall be tested in both human and simian cells.

New text

A strategy for testing adventitious viruses in vaccines must be developed based on a risk assessment. Relevant culture methods and/or specific molecular biology or broad molecular methods should be part of the overall testing package with the agreement of the NRA. In vivo tests may only be used if the risk assessment indicates that this test provides an additional risk mitigation taking into account the overall testing package.

Original text

Each final lot shall be tested for the absence of abnormal toxicity in mice and guinea-pigs by appropriate tests approved by the national control authority.

New text

Original text

Each final lot shall be tested for the absence of abnormal toxicity in mice and guinea-pigs by appropriate tests approved by the national control authority.

New text

Original text

Each final lot shall be tested for the absence of abnormal toxicity in mice and guinea-pigs by appropriate tests approved by the national control authority.

New text

Original text

The cells of the MWCB are suitable if at least 80% of the inoculated animals and eggs remain healthy and survive the observation period, and none of the animals or eggs shows evidence of the presence in the cells of any extraneous agent.
Tests in animals. The following groups of animals shall be inoculated with MWCB cells by the intranluscular route: at least 10' cells being dividedequally between the animals in each group:
-two litters of suckling mice, totalling at least ten animals, less than 24 hours old
- ten adult mice weighing 15-20 g
- five guinea-pigs weighing 3 50-45 0 g
- five rabbits weighing 1.5-2.5 kg.
The animals shall be observed for at least four weeks. Any animals that are sick or show any abnormality shall be investigated to establish the cause of illness. The test is not valid unless at least 80% of the animals remain healthy and survive the observation period.
In some countries, the suckling and adult mice are also inoculated by the intracerebral route.

New text

Original text

The cells of the MWCB shall be shown to be free from potential tumorigenicity by appropriate animal tests, including positive controls, approved by the national control authority. Suitable tests in immunosuppressed animals are as follows. Approximately 10^6 cells obtained from cultures at the same passage levels as those to be used for vaccine production are injected into: newborn mice or hamsters treated with antilymphocyte serum; or athymic mice (nude nulnu genotype); or thymectomized, irradiated mice with reconstituted bone marrow (T-B+). Some of the same group of animals should be inoculated with a similar dose of HeLa or KB cells as positive controls. The animals should be observed for not less than three weeks. Other tests in animals treated with immunosuppressive agents and with equal sensitivity to neoplastic cells may also be used. The test is valid if the positive control animals develop tumours. The cells are suitable for vaccine production if at least 80% of inoculated animals remain healthy and survive the observation period, and none of the animals shows evidence of tumour formation attributable to the cells. "

New text

Original text

At the end of the observation period an amount equivalent to 25% of the control cell-culture suspension shall be tested for the presence of haemadsorbing viruses. If the erythrocytes have been stored, the duration of storage should not have exceeded 7 days, and the temperature of storage shall have been in the range of 2–8 °C.

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

A sample removed from each virus harvest should be tested for virus content by intracerebral inoculation of mice or by plaque assay. If a plaque assay is used a laboratory shall eatablish a correlation between the plaque assay and the mouse LD50 assay.

New text

A sample removed from each virus harvest should be tested for virus content by a suitable immunochemical method, preferably using monoclonal antibodies, or by virus neutralisation in cell cultures.

Original text

Each single cell harvest shall be tested for virus inactivation. The test shall be performed prior to pooling with a sample of undiluted virus suspension by a method approved by the national control authority.
The test sample should correspond to at least 20 human doses of the final bulk. The total volume of the test sample should be inoculated into a primary chicken fibroblast cell culture, or a culture of any other cells with no less suseptability to the virus than chicken embryo cells, and inclubated at a temperature of 37± 1 °C for a period of 14 days. A cell culture sheet of not less than 3cm2 should be used for each 1ml of the test material. No cytopatjic change should be detected at the end of the incubation period. At the completion of the observation period, the culture fluid shoudl be collected and inoculated at a dose of 0.03ml into at least 10 mice of about 4weeks of age. The animals should be observed for 14 days. The single harvest passes the test if the product has been shown to be free from residual live virus.

New text

Each single cell harvest shall be tested for virus inactivation. The test shall be performed prior to pooling with a sample of undiluted virus suspension by a method approved by the NRA. Inoculate a quantity of the inactivated harvest equivalent to not less than 10 human doses of vaccine in the final lot into a suitable cell culture, shown to be sensitive to tick-borne encephalitis virus, with not less than 3 cm2 of cell sheet per millilitre of inoculum. Incubate at 37 ± 1 °C for 14 days. No cytopathic effect is detected at the end of the incubation period. Collect the culture fluid and examine for the presence of infective tick-borne encephalitis virus by a validated in vitro method. If a suitable in vitro method is not available or appropriately validated a suitable in vivo assay may be used with approval of the NRA.