Cell Line Development

Plant-Cell Cultures and Cell Lines for Recombinant Protein Expression

Cell cultures derived from mammalian and bacterial cell lines are the conventional production systems in bioprocessing. But they also have their limitations. Media for mammalian cultures in particular are notoriously expensive, and traditional cell cultures can be highly sensitive to growing conditions. During the late 1980s and into the 1990s, plants and plant-derived cell cultures were introduced as alternative cell-culture systems (1, 2). Although transgenic plants (genetically modified) once looked promising in the early 2000s, the cost and manufacturing complexity…

Direct Analysis of Bioreactor Harvest for Clone Selection and Process Optimization

Therapeutic monoclonal antibodies (MAbs) mostly are manufactured using bioengineered mammalian cells cultured in a bioreactor for two to three weeks. High temperatures and an altered redox environment may compromise the quality of MAbs produced (e.g., fragmentation, truncation), as can the presence of proteases, reductases, and other chemicals released from dead cells. Thus, it would be valuable to establish analytical methods that can help cell culture groups monitor immunoglobulin G (IgG) product integrity in real time during a bioreactor run, especially…

The Upstream Perspective: Taking Efficiency Beyond Cell-Line Development

With 20 years of experience in the biopharmaceutical industry — at Genentech, Applied Biosystems, Cell Genesys, Cellerant Therapeutics, and Bayer — Yuval Shimoni has written frequently for BioProcess International on a number of production topics. Those have ranged from process improvements and bioreactor scale-down validation, to raw materials management, to addressing variability and virus contamination events. For this featured report, we discussed hardware and instrumentation, quality by design (QbD) and related approaches, and other strategies that can take expediting upstream…

Anticipating Cell-Line Challenges to Drive CMC Readiness

Development of a safe and high-quality Chinese hamster ovary (CHO) cell line is of paramount importance for the chemistry, manufacturing, and controls (CMC) portion of studies that support investigational new drug (IND) applications (1, 2). Desirable attributes of a CHO cell line include its ability to produce high titers of biotherapeutic proteins facilitate quick recoveries and selection processes maintain phenotypic and genetic stability throughout in-vitro aging of a culture. A CHO cell line also should be scalable to high-capacity culture…

Ask the Expert: Accelerating Timelines By Integrating Cell-Line Development and Manufacturing

In a 31 October 2019 “Ask the Expert” presentation, Nicole Wakes (group leader of Abzena’s cell-line development team) observed that drug sponsors often outsource their early upstream activities to a few different contract research organizations (CROs). But that strategy can thwart short timelines and introduce regulatory and financial risks. Wakes described Abzena’s upstream approach, illustrating how partnering with a single, multicompetent CRO from cell line construction through manufacture can streamline workflows. Integrating cell line development and manufacturing in this way…

Cell Viability in Bioprocesses: Making a Case for Reevaluation

Trypan blue dye exclusion first was proposed as a means of measuring mammalian cell damages over a century ago in 1917 (1). Despite extensive documentation of its limitations (2), it remains the “gold standard” method of measuring cell viability in common use today. But can this method truly measure viability? And how do we define cell viability, for that matter? Those fundamental questions are linked to whether we refer to cells as “alive” or “dead” in the context of bioprocessing…

Introduction: Reporting from the Frontiers of Cell Line Engineering at BPI Europe and BPI West

Every biomanufacturing process begins with transfection of recombinant genes into pools of cells — followed by a succession of screenings from which will emerge (ideally) a single progenitor cell of the new production cell line. Cast aside will be those cells that do not uptake the correct genetic material, those incapable of thriving in bioprocess conditions, those that fail to produce recombinant protein at relevant levels, and those without demonstrated clonality and relative genetic stability. Over the past several years,…

Creating Novel Cell Lines By Genome Editing: Simplifying Cell-Based Assays and Improving Production of Biomolecules

Cultured cell lines have a diverse range of applications. They are used broadly by cell biologists, clinicians, tissue engineers, biotechnology scientists, and bioengineers. The most important uses of cell culture are in the cell-based assays and production of biologically active recombinant proteins. In recent years, genome editing has been used widely to study the structure, function, and localization of endogenous proteins in cultured cells. However, applying the same genome editing techniques to cell lines also could improve the propagation of…

Time Is of the Essence: Optimizing Cell Line Development

Cell line development is a critical upstream step for the manufacture of monoclonal antibodies (MAbs). Cells must be engineered to produce a biologic of interest, and the right clone must be selected to deliver material for preclinical and clinical studies. A high-producing cell line is then needed to support clinical studies and, ultimately, commercialization of the therapeutic. Please fill out the form below to read more. Corresponding authors are Aurore Poles, Global Technical and Compliance Expert, and Guillaume Plane, Global…

A Faster Solution to Hybridoma Screening

Monoclonal antibodies are a rapidly growing class of therapeutics and are used in multiple clinical indications. This highly competitive landscape necessitates the need for rapidly developing next generation antibodies against new challenging targets with improved mechanism of action and pharmacokinetics. Current antibody screening tools such as ELISA, only report on binding to one antigen target at a time; hence these assays are time consuming and require large amounts of target protein. This case study highlights how the Intellicyt® iQue3 system…