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Biotechnology Services
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Cell Bank, Additive and Bulk Product Testing
Covance offers extensive capabilities in the characterization and biosafety testing of cell banks used in the production of recombinant DNA products, monoclonal antibodies, viral vectors and other biopharmaceuticals. We have over 12 years of experience in performing regulatory compliant programs that have been accepted by the FDA and European regulatory bodies. You can rely on our scientific and regulatory knowledge to help you efficiently develop your products to meet all the latest regulatory requirements worldwide.
Topics discussed below include:
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Cell Bank Biosafety Testing
Regulatory guidelines require that a seed lot system is set up. It consists of frozen cell stocks made up of a Master Cell Bank (MCB) derived from a single cloned cell, a Working Cell Bank (WCB) derived from the MCB and Maximum End of Production Cells (MEPC) comprising cells having undergone a number of population doublings beyond the end of the production process.
Covance can provide GMP compliant cell banking, cell bank storage and the full range of associated testing (see below) to provide our clients with fully qualified cell banks. Where the presence of specific viruses is of particular concern, the general screening assays described below are supplemented with specific assays employing a variety of techniques including immunoprobing, PCR, nested PCR and RT-PCR. |
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General Cell Bank Screening — eukaryotic and prokaryotic
- Authenticity assays, including direct sequencing, high-resolution restriction mapping, isoenzyme analysis, and karyological analysis
- Assays for murine retroviruses, including co-cultivation assay; reverse transcriptase and DNA polymerase single timepoint assay with and without concentration by centrifugation; transmission electron microscopic analysis of cells; enumeration of virus-like particles in cell supernatant by electron microscopic analysis
- Electron microscopic analysis
- Screening assays for adventitious virus, including in vitro evaluation of adventitious viruses in cell cultures; in vivo evaluation of adventitious viruses
- Assays for specific adventitious agents, including mouse antibody production (MAP) assay; immunoprobe assays for specific viruses and PCR
- Mycoplasma and sterility assays, including mycoplasma assay (meeting the requirements of the Points to Consider in the Characterization of Cell Lines, 1993); sterility test (Code of Federal Regulations) or the European Pharmacopoeia
- Testing of insect cell-based production systems for contamination of tissue culture reagents, intrinsic contamination of insect culture
- Spiroplasma testing of insect cells.
Note: Although the above listing is focused on eukaryotic systems, we also provide cell bank screening for prokaryotic systems. This testing includes identification, purity analysis, genetic stability and restriction mapping. |
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PCR Testing
A number of viruses are non-cultivable, latent or of particular concern. Therefore, specific PCR-based assays are necessary to detect these potentially contaminating viruses. In addition to cell bank testing, it is necessary to screen raw materials used in the development and manufacture of cell banks (e.g. trypsin and FCS) to ensure that viral contaminants are not added during the production process.
We have developed a number of specific PCR assays for a wide range of human and mammalian viruses. Quantitative PCR can be used in the detection of retroviruses using a PERT assay. This is extremely sensitive and should be used with caution but is available where circumstances dictate.
Please contact us for an up-to-date list of available assays. If we do not have an assay currently available to meet your requirements then we can rapidly custom-design and develop a robust and sensitive assay. Features of our PCR testing service include:
- Employment of extraction methods appropriate to cell or raw materials
- Range of viral-specific PCR assays available (e.g. HIV, HTLV, HHV, EB, PPV, Bovine Polyoma Virus, MVM, etc.)
- Nested primer sets can be employed to increase assay sensitivity
- More than one set of primers to a particular viral genome can be used to increase validity of data
- PCR results can be confirmed by Southern blot hybridisation
- Fully audited final report for regulatory submission
- Primer design incorporating database searching to ensure specificity.
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Raw Material Testing
Raw materials used in the production of biopharmaceuticals are known to potentially harbor a range of adventitious agents, and hence it is important to screen them for a variety of potential contaminants. The number and type of tests required for each component is determined on a case-by-case basis and applied as dictated by circumstances.
To conform to the CFR 9CFR113.28, each lot of bovine serum to be used in manufacture must be subject to sterility, mycoplasma and in vitro adventitious virus assay (the latter utilising immuno and PCR endpoints for specific bovine viruses). |
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Batch and Lot Release Testing on Bulk Harvest Material
It is important to monitor the bulk harvest of a product in case of a breakdown in GMP. Assays are performed on a case-by-case basis depending on the product type. |
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