GMP Cell & Virus Bank Characterization

Safety testing for GMP cell and virus banks

The safety testing of cell or virus banks at the level of the Master, Working and End of Production Cell Bank is a key element for cell- or virus-derived biopharmaceutical products. Characterization tests aim to confirm identity, genetic stability and purity of the cell or virus bank.

The battery of tests to apply is often adapted depending on the type of cell line or virus seed under development. Virus banks might require neutralization procedure prior to testing. ViruSure’s experts are happy to give best advice on the testing requirements requested by regulatory authorities

What our Clients Say

Polina Kaidun , Head of the cell banking group, BIOCAD (Russia)

European quality, professional approach and efficiency are the main advantages of ViruSure, with which we have been cooperating for several years. The employees are always very helpful and kind, give quick answers to inquiries and are ready for constructive dialogue in disputable situations. We wish the company sustainable development and prosperity.

Recommended Assays for Characterization of Recombinant Cell Banks

The majority of the tests are done at the Master Cell Bank (MCB) and End of Production Cell Bank (EPCB) level. Working Cell Bank (WCB) will be done with a reduced testing package.

Identity Assay
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Identity Testing via QPCR – Cytochrome C Oxidase Gene Amplification

Assessment of the identity and purity of the cell bank lines is a critical part of the quality evaluation. Species like cow, hamster, pig, mouse and human are covered by the qPCR assays using primer sets designed on the Cytochrome C Oxidase gene.
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Purity Assays
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Microbial Contaminants

Sterility according to EP/USP/JP

Sterility Testing is performed to detect the presence of microbiological contaminations and is carried out under aseptic conditions using the Direct Transfer method. An Evaluation of each sample matrix is required once in the course of a Method Suitability to demonstrate that the sample isn’t bacteriostatic or fungistatic. The sample is tested in two different broth to cover the detection of possible anaerobic bacteria as well as aerobic bacteria and fungi contaminations.
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Mycoplasma according to EP/JP

The Mycoplasma qPCR assays are designed on the 16S rRNA gene and are performed according to the European and Japanese Pharmacopoeia.
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Mycoplasma according to FDA PTC

Mycoplasma Testing is traditionally performed as culture method combined with an indicator cell method to detect the presence of mycoplasma species but is now adays replaced by qPCR testing. The Culture method consist of culturing the sample on agar media directly, as well as inoculation of broth media with multiple subculture on agar media. The indicator cell culture method is most common using Vero cells which are stained with a fluorescent dye to show possible contaminations. Mycoplasmastasis testing is performed at least once for each sample matrix to detect any presence of inhibitory substances which might affect the assay.
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Adventitious Viruses – General Virus Screening

28-day in vitro assay for the presence of viral contamination using 3 cell lines

The in vitro assay uses a selection of cell lines with a proven history in the detection of a wide range of potential virus contaminants. It is used both in the testing and characterization of recombinant cell banks, as well as in the testing of raw materials (e.g. bovine serum). Cell lines include Vero, MRC-5, HeLa and CHO. Cell lines include Vero, MRC-5, HeLa, and CHO.
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In vivo assay for adventitious agents in suckling and adult mice, embryonated eggs

The in vivo adventitious agent test in suckling mice, adult mice, embryonated eggs or guinea pigs investigate the presence of virus contaminations that cannot be detected in cell culture.
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Retrovirus Tests
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Transmission electron microscopy (TEM)

TEM allows visualization of virus particles, particle size, morphology, and location.
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Retrovirus infectivity test; extended XC plaque assay and extended S+L- focus assay

Many cells (especially cells of rodent origin) endogenously express retrovirus particles, or contamination may be introduced into the cell line exogenously. In vitro assays are used to detect the presence of replication competent retroviruses in recombinant cell banks or bulk harvests. Cell lines include PG4 S+L-, Mus Dunni, XC.
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Reverse transcriptase assay using the PERT assay (FPERT)

Fluorescent Product Enhanced RT Assay is used when highly sensitive qualitative fluorescent results are required for testing levels of retrovirus in vaccine or recombinant bulk.
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Other Viruses – Specific Virus Tests
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Mouse antibody production test (MAP, only applicable to rodent cell lines)

The MAP (mouse antibody production) test investigates viral contamination in the blood after a defined observation period by the analysis of antibody titres towards a defined panel of viral antigens.
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Hamster antibody production test (HAP, only applicable to rodent cell lines)

The HAP (hamster antibody production) test investigates viral contamination in the blood after a defined observation period by the analysis of antibody titres towards a defined panel of viral antigens.
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Bovine viruses *

Bovine viruses can potentially contaminate biologics and biotechnology products due to exposure to bovine derived media components (e.g. FBS). A package of qPCR tests for common bovine viruses is offered.
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Porcine viruses *

Porcine viruses can potentially contaminate biologics and biotechnology products due to exposure to porcine derived reagents (e.g. trypsin). A package of qPCR tests for common porcine viruses is offered.
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Human viruses (recommended for human-produced cells) *

Tests for human viruses is recommend for human producer cells or for a new production system which has been handled in a R&D environment to establish the cells and lay down a research bank. A package of qPCR for common human viruses is offered.
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Simian viruses *

Tests for simian viruses is recommend for simian producer cells. A package of qPCR tests for common simian viruses is offered.
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Avian viruses *

For biopharmaceuticals products produced in avian cells testing for specific viruses is recommend. A package of qPCR for common avian viruses is offered.
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Insect viruses *

For cell lines of insect origin, tests for specific viruses relevant to the species of origin of the insect cells and for arboviruses (arthropod-borne viruses) are recommended. A package of qPCR for common insect viruses is offered.
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* Testing requirement will depend on the species of the seed cells, on their history and to which raw materials they have been exposed.

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