BHK-21 Cell Line Clonality Assessment

BHK-21-based production are the dominant technologies for veterinary vaccines and recombinant protein production. These cell lines can be easily adapted to grow in suspension cultures.

In recent years proof of clonality has become a focus. Many companies have received comments back on this topic during the IND review process of their biological products. Clonality is thought to minimize the heterogeneity of cell banks and thus allow for consistent manufacture a product. Here, Creative Bioarray has established a high-throughput clonality analysis (FISH) strategy to fast-track your stable cell line development and accelerate subsequent IND review.

There are regulatory considerations regarding the clonal derivation of the cell line used to create the master cell bank (MCB). Though no one single regulatory guidance speaks definitively to the need of having a demonstrably "clonally derived" cell line, multiple guidance documents allude to this general expectation and underscore it as a basic expectation. These include, but are not limited to:

• ICH Q5D Derivation and Characterization of Cell Substrates Used for Production of Biotechnological/Biological Products, "For recombinant products, the cell substrate is the transfected cell containing the desired sequences which has been cloned from a single cell progenitor"
• US FDA Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use notes that "The MCB is defined as a collection of cells of uniform composition derived from a single tissue or cell",
• EMA/CHMP guidance notes "The cell substrate to be used for the production of the monoclonal antibodies should be a stable and continuous monoclonal cell line that has been developed by means of recombinant DNA and/or other suitable technologies".

Pattern 1 (200/200 cells) Telomeric region of chr.8

Creative Bioarray's BHK-21 Cell Line Clonality Assessment Service has the following Features:

• Detection of each individual integration site
• Analyze 100-200 cells per sample
• Rapid 4-week turnaround time, including addition reporting
• High accuracy and sensitively
• Competitive pricing

Creative Bioarray offers BHK-21 Cell Line Clonality Assessment Service for your project as follows:

• Probe design
• Probe synthesis
• Chromosome spread preparation
• FISH on chromosome spreads Imaging
• Data analysis (karyotyping and integration pattern analysis)

Quotation and ordering

Our customer service representatives are available 24hr a day! We thank you for considering Creative Bioarray as your BHK-21 Cell Line Clonality Assessment Service partner.

References

1. Frye, Christopher, et al. "Industry view on the relative importance of "clonality" of biopharmaceutical-producing cell lines." Biologicals 44.2 (2016): 117-122.
2. Wurm, Florian M., and Maria João Wurm. "Cloning of CHO cells, productivity and genetic stability—a discussion." Processes 5.2 (2017): 20.
3. ICH Q5D Derivation and characterisation of cell substrates used for production of biotechnological/biological products (CPMP/ICH/294/95)
4. 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.
5. 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).
6. Wu P, Hartman T, Almond L, Stevens J, Thrift J, Ojha J, Alves C, Shaw D, Laird MW, Emmins R, Zhu Y, Liu R, Du Z, Koehler R, Jostock T, Anderson K, Campbell C, Clarke H. Advancing Biologics Development Programs with Legacy Cell Lines: Advantages and Limitations of Genetic Testing for Addressing Clonality Concerns Prior to Availability of Late Stage Process and Product Consistency Data. PDA J Pharm Sci Technol. 2020 Mar-Apr;74(2):264-274. doi: 10.5731/pdajpst.2018.009316. Epub 2019 Sep 13. PMID: 31519780.