If you have a cell line (not subcloned) with variable growth properties, how would you validate the process from cracking of the vial through isolation of product?
Hello Maureen,
The validation process would include verifying the identity of the cell line by not only testing the cell line but also documenting the history of the cell line. Once enough documentation on the history and characteristics of the cell line is collected, you may use various methods to authenticate the cell identity such as morphology check, karyotyping and gene sequencing. You may also use phenotypic method such as growth curve analysis to validate the cell line. Isoenzyme analysis will help to identify species of origin and detect cross contamination. STR analysis can help detect contamination as well as genetic drift in cell culture.
In cell line validation process, you need to consider purity and sterility as well. Cross contamination may compromise product yield, quality and safety. Adventitious virus testing should be done. For example, for human cell line, testing for HIV and other viruses that infect human cells should be done. Mycoplasma contamination should also be tested as it can cause decreased cell growth and protein production.
Finally, in the validation process, one needs to assess the cell line's ability to consistently produce high-quality protein. This may be done by checking the mRNA levels by qPCR and the protein by Western Blot analysis by using a specific antibody to ensure that the protein is the right protein that you are interested in. Also, one could design a bioassay to test the bioactivity of the protein of interest.
You may want to refer to chapter 36 of the book attached below.
https://doi.org/10.3109/9781420019797-37
Best.
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