Genetic Engineering & Biotechnology News

JUL 2018

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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16 | JULY 2018 | Genetic Engineering & Biotechnology News | GENengnews.com See Cell Line Validation on page 18 Failing to correctly validate cell lines used to produce therapeutic proteins can have substantial financial repercus- sions for biopharmaceutical manufacturers and, in extreme scenarios, pose safety risks for patients. In the manufacturing game, validation means verifying not only identity but also purity, sterility, and functionality in ways that meet regula- tory agency standards. Confirming Identity Confirming cell identity doesn't actually begin at the lab bench. "Documentation of the history of the cell line is just as important as the testing of that cell line," explains Michael J. Hantman, Ph.D., associate director, methods development and validation, at Charles River Laboratories. "If you can't say where the cell line came from—that's going to be a big red flag for regulatory agencies." Once the manufacturer has collected adequate documen- tation on the history and characteristics of the cell line, it can apply a variety of techniques to authenticate cell iden- tity—ranging from simple morphology checks, to classic karyotyping analyses, to modern gene sequencing methods. Phenotypic methods, such as cell morphology and growth curve analysis, are perhaps the simplest methods used to validate cell lines. The American Type Culture Collection (ATCC) recommends frequent, brief observations of cell morphology, which can change depending on culture density, differentiation state, cell health, and media composition. Un- usual morphology can indicate underlying issues—including contamination with another cell line or microorganism. Sim- ilarly, sudden changes in growth can also indicate a problem; it's important to monitor cell growth for consistency using a growth curve analysis. Isoenzyme analysis provides another relatively rapid and inexpensive method to identify species of origin and detect cross-contamination. Different isoforms ex- ist for a number of intracellular enzymes, and isoenzyme analysis uses electrophoretic banding to reveal subtle differences in the structure and motility of these variants— creating distribution patterns unique to dif- ferent species. One limitation is that isoen- zyme analysis cannot distinguish between cell lines from the same species. Regarding karyotyping, according to Dr. Hantman, it's "a classic method but really informative." Scientists can use the number and appearance of chromosomes to confirm species of origin and identify human cell lines with unusual chromosome patterns sometimes observed in lines researchers have used for a long time. Turning to gene sequencing, advances in the technology have made this contem- porary technique more accessible and af- fordable—leading to its adoption in a wide array of applications, including cell line characterization. The extent of sequencing can range from a single gene to the whole genome. For species-of-origin analysis, DNA bar- coding is a popular technique. Referred to as the "Barcode of Life," the highly conserved region in the mitochondrial cytochrome oxi- dase subunit I (COI) gene provides a short Bioprocessing Cell Line Validation in Biomanufacturing Meghaan M. Ferreira, Ph.D. Cell line validation has been under a bright spotlight since evidence emerged of widespread misidentification and contamination of cell lines, including those used in preclinical research and cited in thousands of papers. And preclinical research is not the only arena where misidentified or contaminated cell lines can wreak havoc and undermine years of work. A multimodal approach provides greater confidence that a particular cell line is authentic and will provide the functionality needed for efficiency, productivity, and safety. The AVATAR ™ Cell Control System More Cells | Better Cells | Faster | On Target Gain control over your primary cells to achieve: - Rapid expansion in less time (CD8+ T-cells at half the time to target cell number) - Increased viability post transfection/transduction - Targeted naive and memory cells without the need for activating antibodies (CD3/CD28) - Better functional response and characterization in primary tumor cells - Shortened vein-to-vein time www.xcellbio.com ©2018 Xcell Biosciences, Inc. All Rights Reserved. The trademarks used herein are the property of Xcell Biosciences, Inc. or their respective owners. FOR RESEARCH USE ONLY. NOT FOR IN VITRO DIAGNOSTIC USE. Mimic the Human Microenvironment (HME™)

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