Determining the Right GMP Safety Testing Plan for Your Biologic

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BI-000686

The risk of viral and microbial contamination is a feature common to all biological products derived from cell lines. In order to comply with Regulatory Agencies like the FDA and EMEA, all cell banks, raw materials of animal or human origin, bulk harvest samples, and final drug products are subjected to various safety tests to ensure that they are free of any possible contaminants. Such contamination can arise from the source cell lines themselves (cell substrates) or during the production process, and can have serious clinical consequences. 

Testing Requirements

The testing required for different types of samples will depend on the product itself, the regulatory guidelines being followed, and what possible contaminants could have been introduced from the raw materials and manufacturing process. Examples of testing panels are below. 

 Mammalian Cell Substrate Testing Panels1
    Master Cell Bank Working Cell Bank End of Production/Cells at Limit Bulk Harvest
Microbial Contamination

Sterility/Bioburden

X X X X
Mycoplasma X X X X
Mycobacterium X      
Spiroplasma X      
Cell Line Identity DNA Barcoding X X X  
DNA Fingerprinting/STR X   X  
Karyology X   X  
Genetic Stability DNA Sequencing X   Optional  
Gene Copy Number by qPCR X   X  
Virus Testing Retroviruses: Reverse transcriptase and retroviral infectivity assays X   X X
In vitro adventitious viruses X X X X
In vivo adventitious viruses X   X X
Test for specific viruses (e.g., bovine, porcine, human, minute virus of mice) X     X
Mouse, hamster, and rat antibody production assays (MAP, RAP, and HAP) X      
Transmission electron microscopy X   X X
Tumorigenicity/Oncogenicity     X  

1Novel molecular methods based on next-generation sequencing technology can also be offered to supplement the test program.

Our Ballina, Ireland facility has recently added to their portfolio of available tests to detect viral and microbial agents with the qualification and implementation of in vitro AVA and mycoplasma testing capabilities. The addition of these assays compliments the in vivo methods already in place to detect viral agents (i.e., in vivo biosafety and mouse, hamster, and rat antibody production assays). Having both in vitro and in vivo, and microbiological capabilities (e.g., sterility, bioburden) at the Ballina site allows us to offer a complete package of safety testing at a single location, giving you the ability to streamline your submission process and get results quickly. Along with the packages, Ballina is also offering a sample pickup service for our clients in Ireland.

Ballina is now able to offer the following testing for your samples:

Microbiological Tests

  • Sterility testing: clean room or isolator (Ph.Eur. 2.6.1., USP<71>, ISO11737-2)
  • Bioburden (ISO 11737-1)
  • Endotoxin: gel clot, kinetic turbidimetric, and kinetic chromogenic LAL (Ph.Eur. 2.6.14., USP<85>)

In Vivo Assays

  • In vivo test for adventitious agents (EP 2.6.16., US FDA Guidance for Industry 2010 and ICH Q5A or combined method)
  • MAP/HAP/RAP

In Vitro Assays

  • In vitro test for adventitious agents
    • Positive control viruses available: parainfluenza-3, minute virus of mice, sindbis virus
    • Detector cell lines available: MRC-5, Vero, 324K, CHO-K1Readouts, cytopathic effects, hemadsorption, and hemagglutination

Mycoplasma Testing

  • Indicator cell assay (USP, EP, JP, and FDA PTC 1993)
    • Vero cells are cultured in the presence of the test product and then stained with a DNA-binding fluorescent dye
    • Only nuclear DNA staining is observed in cells that are free of mycoplasmas
    • Mycoplasma are detected by their characteristics particulate of filamentous pattern of fluorescence on the cell surface 
  • Culture method: Agar and liquid media (USP, EP, JP, 21 CFR 610, and FDA PTC 1993)
    • Test products are inoculated directly onto agar plates as well as into liquid medium - both support the growth of a broad range of mycoplasma species
    • Agar plates are inspected for the presence of mycoplasma colonies

Additional supplementary testing is available at our facility in Erkrath, Germany:

  • Retrovirus assays
  • Electron microscopy
  • Bovine and porcine assays

If you are developing a biological product and ready to perform safety testing, Ballina offers a complete package to make the process easier and meet your unique testing needs. For more information and/or to schedule a consultation, contact us at [email protected].