HEK293 Cell Line Genetic Stability (MCB, WCB, EOP) Testing

Over the past several decades, numerous therapeutic recombinant proteins (e.g., monoclonal antibodies, bi-specific monoclonal antibodies, growth factors, cytokines, hormones, enzymes, and vaccines) have been approved by regulatory authorities, and many more are currently undergoing clinical development. HEK293 cells are the predominant hosts for expression of recombinant proteins and are used for stable expression of proteins.

Genetic stability testing is an important component of HEK293 production cell bank characterization and is a regulatory requirement. During subsequent HEK293 cell culture, genomic events such as deletion, rearrangement, and point mutation may occur and result in an altered cell phenotype or gene expression profile. The instability of the HEK293 production cell line is of great concern because it may negatively affect product integrity, thus posing a risk to the patients. Creative Bioarray offers HEK293 Cell Line Genetic Stability (MCB, WCB, EOP) Testing to characterize Master Cell Banks (MCB), Working Cell Banks (WCB), and End of Production Cells (EPC) from HEK293 cell cultures.

According to ICH Q5B and Q5D guidelines, regulating agencies request that cell substrates must be analyzed at two time points (i.e., on the level of the Master Cell Bank and on the level of the End of Production cells) to show their genetic stability. Typical assays include those intended to confirm the integrity of the product transcript, the genome structure at integration site, and the ratio of insert gene copy number.

Genetic Stability Testing Methods

Creative Bioarray applies the following methods to determine the genetic stability test of recombinant HEK293 production cell lines:

• Transgene insertion copy number analyzed by qPCR and/or Locus Amplification & Sequencing
• Structural analysis (insertion sites/restriction analysis) by qPCR and/or Locus Amplification & Sequencing
• Transgene integrity pattern analysis by FISH
• DNA and RNA sequencing

The HEK293 Cell Line Genetic Stability (MCB, WCB, EOP) Testing Features:

• Genetic stability analysis across various generations (e.g., MCB, WCB, EOP)
• In accordance with ICH Q5B and Q5D guidelines
• Final conclusions on stability of analyzed cell bank(s)
• Reporting meeting regulatory requirements
• Pre-finalization report review included
• Fast turnaround time

Quotation and ordering

Our customer service representatives are available 24hr a day! We thank you for considering Creative Bioarray as your HEK293 Cell Line Genetic Stability (MCB, WCB, EOP) Testing Service partner.

References

1. ICH Q5B, Quality of Biotechnological Products: Analysis of the Expression Construct in Cells used for Production of R-DNA Derived Protein Products, Step 4 version (1995).
2. Wurm, Florian M., and Maria João Wurm. "Cloning of CHO cells, productivity and genetic stability—a discussion." Processes 5.2 (2017): 20.
3. Plavsic, Mark. "Q5D derivation and characterization of cell substrates used for production of biotechnological/biological products." ICH Quality Guidelines (2017): 375-393.
4. Galbraith, Daniel. "ICH Q5A: Viral Safety of Biotechnology Products." ICH Quality Guidelines: An Implementation Guide (2017): 311-335.
5. Krebs, Lara E., et al. "Effective and efficient characterization of Chinese hamster ovary production cell lines using automated intracellular staining and statistical modeling." Biotechnology Progress 34.3 (2018): 570-583.
6. Welch, J. (2017). Tilting at clones: A regulatory perspective on the importance of "Clonality" of mammalian cell banks. CDER/OPQ/OBP/DBRRIV April 24, 2017.
7. Paul Wu, et al. "Tools and methods for providing assurance of clonality for legacy cell lines" in "Cell Culture Engineering XVI", A. Robinson, PhD, Tulane University R. Venkat, PhD, MedImmune E. Schaefer, ScD, J&J Janssen Eds, ECI Symposium Series, (2018).
8. Frye, Christopher, et al. "Industry view on the relative importance of "clonality" of biopharmaceutical-producing cell lines." Biologicals 44.2 (2016): 117-122.
9. Food and Drug Administration, Center for Biologics Evaluation and Research Food and Drug Administration. Supplement to the Points to Consider in the Production and Testing of New Drugs and Biologic& Produced by Recombinant DNA Technology: Nucleic Acid Characterization and Genetic Stability. 1992
10. Ph Eur, Monographs: Medicinal and Pharmaceutical Substances, Products of Recombinant DNA Technology. 2016
11. ICH Topic Q 5 B, Quality of Biotechnological Products: Analysis of the expression construct in cell lines used for production of rDNA derived protein products. 1996
12. Hughes ED, Qu YY, Genik SJ, Lyons RH, Pacheco CD, Lieberman AP, Samuelson LC, Nasonkin IO, Camper SA, Van Keuren ML, et al. Genetic variation in C57BL/6 ES cell lines and genetic instability in the Bruce4 C57BL/6 ES cell line. Mamm Genome 2007; 18(8):549-58; PMID:17828574; https://doi.org/10.1007/s00335-007-9054-0