China 
Africa 
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Cell Culture Process Validation Including Cell Bank Qualification
Published in James Agalloco, Phil DeSantis, Anthony Grilli, Anthony Pavell, Handbook of Validation in Pharmaceutical Processes, 2021
Anne B. Tolstrup, Steven I. Max, Denis Drapeau, Timothy S. Charlebois
Furthermore, during the past ten years FDA has had an intense focus on “clonality” documentation of the manufacturing cell line, and investigators are expected to provide detailed descriptions on how their production cell lines are generated and to demonstrate a high probability of “clonality” for any novel production cell line (see Rachel Novak, 2017). While biopharmaceutical industry professionals and academic experts have discussed and critically analyzed the rationale and feasibility behind this requirement intensely at conferences and in publications (Frye et al, 2016; Bandyopadhyay et al, 2017), FDA still expects such documentation to be submitted prior to market authorization at the latest. It should be noted that the European Medicines Agency (EMA) has not been following FDA regarding this request for documentation of clonality. The focus on clonality has led to development of new methodologies to support and enable cell cloning and documentation in a faster and more robust manner compared to the traditional (and still regulatory accepted) “two rounds of limited dilution cloning” approach. Examples of newer technologies and instruments which have gained acceptance as methods that can ensure a high probability of clonality include (1) single-cell sorting by flow cytometry instruments combined with imaging of the wells into which the single cells have been deposited; (2) ClonePix driven clone selection combined with one round of limited dilution cloning; and (3) the Beacon Optofluidic methodology, which uses optics/light to move individual cells into separate compartments (termed “pens”) and—after expansion and productivity measurements in the pen—exports individual selected clones for further culture.
Evaluation of different vector design strategies for the expression of recombinant monoclonal antibody in CHO cells
Published in Preparative Biochemistry & Biotechnology, 2018
Hadi Bayat, Saghar Hoseinzadeh, Eśhagh Pourmaleki, Roshanak Ahani, Azam Rahimpour
Single-cell cloning was performed by standard serial dilution cloning. Cells were diluted to the density of 10 cells/ml in Nutrient Mixture F-12 medium supplemented with 15% FBS and nonessential amino acid mixture (Thermo Fisher Scientific, USA). About 100 µl of each dilution was seeded in 96-well plates. Single clones were identified after 2 weeks of incubation and were transferred to 24-well plates when they reached to 70% confluence.