How to Optimize Media and Improve Protein Production with Defined Supplements

Enhancing protein production is a common goal in the biomanufacturing industry. During this Ask the Expert session, we will be discussing the challenges associated with animal origin free media optimization and how defined supplements can provide an increase in productivity and yield. Essential Pharmaceuticals is addressing the challenge of animal origin free media optimization with their Cell-Ess supplement. Use of this product at 1% concentration resulted in a 37% increase in productivity. When using the supplement as a feed it resulted in in a 25% increase in yield and an extension of peak protein production with functional protein increase and desired glycosylation achieved. Results suggest that an increase in protein production may not necessarily require a change in the metabolic state of the cells.

Who should visit?

  • Those in early development of bioproduction of proteins in eukaryotic cells
  • Those in media optimization or yield optimization for scaled bioproduction of proteins
  • Department Directors, Program Directors, Project leaders, involved in optimization of large scale bioproduction of proteins.

Types of Questions?

  • What benefits can be gained by using a defined animal origin free supplement in bioproduction?
  • Can peak protein yield be extended?
  • How does adding a boost or feed impact the metabolic profile?
  • How can supplements be used in impact large scale bioproduction?
  • Can one increase protein yield without increasing biomass, i.e. making cells be more efficient at producing the engineered protein of interest?

This Ask the Expert session is sponsored by Essential Pharmaceuticals and is hosted by Dr. Adam Elhofy, Ph.D., CSO. Dr. Elhofy developed the core technology for the Ess line of products and aided in creating patents around novel uses of materials.  Dr. Elhofy has over 14 years of scientific research experience in the areas of immunology, neuroscience, and oncology.  He was funded by both the National Institutes of Health and the Multiple Sclerosis society as an investigator at Northwestern University Medical School. His doctoral research won him the award of the Top 5 trainee scientists by the American Association of Immunologists. Dr. Elhofy has 14 scientific publications in peer reviewed journals. He has played a variety of roles with start-up biotech companies ranging from Principal Investigator to Director of Corporate Development.

For more information, please see “Achieving control and reproducibility of cell culture by eliminating serum.”


Question 1

With titers at 3g/l or in most cases even better, we aren’t necessarily looking for improved titer. Are there other benefits to be gained?

This is great question. I will try to address this from 2 different perspectives.

The first way to think about a benefit is through logistics. When we have used Cell-Ess as a feed, we found the increased output occurred early. We reached the max yield of the control group 2 days earlier than in the control. In this case you could have the benefit of ending a run 2 days earlier.

The other half to consider is protein quality. Quality is measured through many assays. The quality of a protein is driven by the endoplasmic reticulum and Golgi apparatus. Cell-Ess may support better health of both of these organelles. We are currently running assays to access what, if any, are the effects of Cell Ess on protein quality.

Question 2

How does your product impact downstream purification?

This is an important question. We wanted to understand if Cell-Ess could affect downstream processing. To date we have not seen any effect in HPLC purification or Protein A purification. At this point we do not anticipate any effect of Cell-Ess on the downstream purification process.

Question 3

Is there any impact to the metabolic profile?

This is an interesting question. I believe it is asking what can happen with a new additive or increasing protein production. Cell Ess while increasing protein production does not alter the metabolic profile of many of the common metabolites. As an example glutamate, lactate, and ammonia are unchanged with the addition of Cell Ess.

Question 4

What cell types have you used your Cell ess with?

Cell Ess has been used with a variety of cell types. Cell Ess has been used in cell lines and primary cells. In cell lines Cell Ess has been used a549, HEK 293, VERO, CHOk1, and many other cancer cell lines both adherent and suspension type of cells. For cell lines there is a adaptation that is required and possible optimization required. For example there are some cancer models using cell lines that are hormonally dependent in those cases it may require the addition of the needed hormone like estrogen.

Cell Ess also works with primary cells. In the case of primary cells there is no need for adaption but there may still be the need for optimization based on the goals of the research. For example there are some cells that require the presence of TGF-b for their function. In those cases you would have to add exogenous TGF-b.

We designed Cell Ess to be used across many cell types for that reason there may be cell specific or lab specific optimization. Once the optimization is determined the results have been consistent and reproducible.

Question 5

When you talk about improving bioproduction, are you adding this supplement to classic or chemically defined production media?

This is an interesting question. We have tried both methods. The most common media used for bioproduction is chemically defined media. In many cased there are internal proprietary formulations. We tested Cell Ess in a chemically defined system as part of an optimization strategy to see if we could improve performance of the system. We found a benefit if we added Cell Ess as either an initial supplement or as feed.

In serum containing systems we adopted the system to serum free and then were able to see similar performance as was in the serum containing media.

Question 6

I am wondering if you know whether this product could replace fbs in vero vaccine manufacturing?

Cell Ess has been used in the adaptation of Vero cells. There are several groups currently testing for viral infection. There are other options instead or going completely animal free. It is possible to go mostly animal free a combination of Cell Ess and 0.1% FBS and maintain most characteristics of the original culture. There are also other supplements like adding BSA that may be beneficial. For Vero cells as with other cells with optimization, Cell Ess has worked well.

Question 7

Can peak protein yield be extended?

The increase in protein production occurs early and stays higher than control.

Question 8

How does the cost compare to FBS? If we are using 3% FBS for a Cell Therapy application. Would it be cost effective to switch to your product, just if you look at a straight switch of supplements not taking into account savings in regulatory resources, etc?

The costs for FBS vary greatly. This question is difficult to answer as the type and source of FBS are key drivers for the cost of FBS. In the past year the price of FBS has been increasing. Cell-Ess currently costs less than many FBS choices that it can replace. The other side of the question is indirect costs. The use of FBS typically is preceded by multiple lot testing which has time and resource costs. In some cases groups will buy entire lots and then there are costs associated with tying up a large sum of money, carrying unused inventory for years and long term material storage in a freezer.

The second part of the question is the interesting, as the one of the primary reasons for switching away from FBS in therapeutics is to ensure consistency in the product. According to a few of the partners we are working with price of having an inconsistent product is more costly than any of the raw material costs. As far as doing pure dollar to dollar cost analysis it is difficult to deliver a meaningful number without knowing the many variables in play.

Question 9

Has this been used in any products under review with FDA?

Cell Ess is currently in trials by several groups. There is a DMF for Cell Ess.

Question 10

Does the product increases the cell specific productivity or increases the total productivity and how does the product impact the glycosylation profile?

This is a great question. We have only measured the target protein increase. We do not know if the Cell Ess also increases endogenous protein production.

We are currently working on the glycosylation profile and should have more data in the next few months.

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