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Maximizing Transient Protein Production
Companies are turning to transient production of proteins to accelerate timelines, gain quick access to protein for early go-no go decisions, delay stable cell line generation, and reduce costs. Expanding the role of transient protein production within the biotherapeutic discovery and development workflow is highly dependent on the ability to supply the required quantities of quality proteins — generally ranging from milligram to multiple grams of protein — in the required timeframes. Thus, increasing productivity and process scalability are aspects of production that must be addressed.
This session is sponsored by MaxCyte
There are a wide variety of factors that influence productivity – – from pre-transfection factors such as cell type, vector design, and cell health to post-transfection factors such as seed densities, media additives, and feed schedules. In addition, researchers must consider the method of transfection which affects both productivity (cell health & transfection efficiency) and process scalability.
Recently, Cell Culture Dish has published several poster and webinar articles discussing the use of MaxCyte’s electroporation-based delivery platform for cell engineering, specifically in the areas of transient and stable protein production. There was a high level of interest from readers regarding CHO-based, gram-level protein production, and so during this week’s Ask the Expert session, we will be joined by Dr. Weili Wang, the Director of Cell Culture at MaxCyte, to answer questions regarding cell engineering and culturing that can maximize transient protein production. Dr. Wang has over 20 years of biopharma industry experience focusing on process development, stable cell line generation, tech transfer and scale up/scale down modeling to support cGMP manufacturing. Prior to joining MaxCyte, Dr. Wang was the upstream manager at Human Genome Sciences, MacroGenics and Amplimmune. Dr. Wang received his Ph. D degree from Texas A&M University, a Master degree from Florida International University and Bachelor degree from East China University of Science & Technology.
Please take advantage of the opportunity to ask our expert a question and participate in a lively discussion on animal origin free media optimization.
Questions & Answers
Would you recommend a media change post transfection or could you tell me how to avoid having to change the media. How long to culture post transfection?After transfection, we typically do not change media, unless we are performing selection for stable pool or clone generation. The high level of cell viability following MaxCyte electroporation allows not only for rapid protein production, but also for longer-term culturing to maximize total protein yield. Thus, the length of post transfection culture depends on the […]» Read More
I liked your article, is the 2.5 g/l average you mentioned pretty reproducible for most CHO lines or do you see lots of variation in the titers?Antibody yields following MaxCyte electroporation using a given expression system and downstream culture conditions is highly reproducible. In terms of the ability to achieve g/L productivity, it is dependent on a variety of factors including the vector, the protein being expressed, and as you inquired, is also dependent on the cell line being used (including […]» Read More
What kind of target types using electroporation, i.e. non-antibody proteins, antibody fragments, fusion proteins?Electroporation can be used to engineer cells to express a range of proteins from secreted proteins, membrane proteins or cytosolic proteins. In terms of protein biotherapeutics, MaxCyte clients have expressed recombinant antigens, full IgGs from multiple species, bi- and tri-specific antibody, FcSv, Fab and Fc, or other fusion proteins. One benefit of electroporation over other […]» Read More
I read your post on host cell choice, but I was wondering is there ever a time when HEK is best or are there certain applications where there is a clear reason to use one over another. Would you ever use a cell line other than CHO or HEK, for instance if the manufacturing cell line was going to be NS0 or SP2/0Hybridoma cells like NS0 and SP2/0 were used as the cell line of choice very early on for the development of therapeutic recombinant protein drugs. CHO cell lines have now been well characterized, have post translational modification profiles closer to human cells than hybridoma cells, and have a regulatory track record of use in manufacturing […]» Read More
Does the quality of the expressed protein decrease if you push transient expression to gram/L titers?The quality of expressed proteins is not necessarily related to high titer. Many factors contribute to protein quality including the molecule itself, transfection conditions, production conditions, and the host cell line. Recently, we observed issues specifically with protein degradation when using the ExpiCHO cell line for several proteins even when using MaxCyte flow electroporation. Interestingly, […]» Read More
Have you produced viral vectors using CHO? Have you seen any differences or benefits over using HEK?In general, we find our clients are using HEK, Vero, or insect cells for producing viral vectors or VLPs. Thus, our research scientists do not have direct experience transfecting CHO cells specifically for viral vector production. We certainly have produced a variety of vaccines and viral vectors in other commonly used cell lines.» Read More
Using your system have you been able to express a variety of antibody types, including ones with kappa or lambda light chains?Yes. Both MaxCyte scientists and our clients have expressed a range of antibodies, antibody-like molecules and Fc fusion proteins from a variety of species including human, rabbit and mouse. In comparison studies with other transfection methods, flow electroporation generally leads to higher titers independent of the type of antibody molecule that is being expressed. GFP […]» Read More
Are differences in cell viability levels from one transfection method to another large enough to actually affect protein titers?Yes, high cell viability is critical for achieving high titers. Of course, it is essential to use cells that exhibit high viability pre-transfection (>95%), but it should also be emphasized that if cell viability is impacted by the transfection process, then cells will also exhibit reduced productivity. We consistently observed that post transfection viability >90% […]» Read More