Media Optimization Can Improve Glycosylation Patterns and Consistency to Impact Protein Efficacy

Sponsored by: Essential Pharmaceuticals
Session ends: December 2nd, 2016, 3:00pm MST
Answers by: Dr. Adam Elhofy, Ph.D., CSO,


When producing a biologic therapeutic, protein quality is a key consideration.  While manufacturing parameters such as high protein yield, protein solubility and protein stability are important, it is also critical that the biologic exhibits the desired therapeutic activity. Post-translational modifications impact efficacy and pharmacokinetics of the biotherapeutic, where improper glycosylation can result in an ineffective drug.  Proteins are commonly post-translationally modified by adding sugar residues in a process called glycosylation.  Half of all human proteins are estimated to be glycosylated.

This session is sponsored by Essential Pharmaceuticals

During this Ask the Expert session, we will be discussing factors that impact glycosylation and tools available to improve the glycoform complexity and increase consistency of glycosylation profiles. Essential Pharmaceuticals is addressing these challenges with their Cell-Ess universal titer boost and optimizer.  Cell-Ess was used as a feed in a Wave bioreactor, resulting in greater than 25% increase in monoclonal antibody titer, which was similar to the titer increases observed in earlier experiments using shake flasks.  In addition to increasing titer, Cell-Ess was shown to significantly increase consistency of glycosylation patterns when used as a feed.  Further, Cell-Ess was shown to increase higher order glycoforms measured by increase in galactosylation.  These new data suggest supplementing with Cell-Ess as a feed increases protein titer while improving or maintaining quality.

Who should visit?

  • Those in process development for bioproduction of proteins in mammalian cells such as CHO 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 factors influence glycosylation patterns?
  • What impact does glycosylation have on efficacy?
  • What factors influence consistency of glycosylation patterns from run-to-run?
  • What are the benefits of the higher order glycoforms?

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 15 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 “Cool Tool – Novel universal titer boost and enhancer improves CHO cell protein production in small bioreactors.

Please take advantage of the opportunity to ask our expert a question and participate in a lively discussion on animal origin free media optimization.

ask the expert

Questions & Answers

Have you looked at any other post translational modifications? lipidation?

We have not specifically looked at other post-translational modifications such as lipidation.  Cell-Ess can improve the lipid membranes including those of the ER and Golgi. Therefore, theoretically there should be a benefit in using Cell-Ess for lipidated proteins, but we have not tested it yet.  Lipidation, similar to glycosylation, is a complicated multistep process where […]» Read More

What is the relationship between glycosylation and aggregation?

There are many reasons for aggregation.  To simplify it, imagine there is the ability to formulate an equation to determine if and how much proteins will aggregate and at what concentration.  The formula would have many inputs.  Glycosylation is one of the many inputs that would determine whether a protein will aggregate.  In the case […]» Read More

I have been seeing articles on the use of supplements, peptones, etc. regulating glycosylation. Do you think it is better to start with a CD media or with a media that already contains some supplements? Why would these supplements be having an impact on glycosylation? Healthier cells?

The optimal way to start would be with a CD media and establish a baseline.  Once you have a baseline for your system, then CD supplements can be added individually to determine whether the profile changes.  If you are not able to change your profile with the CD supplements, then you can move to the […]» Read More

We are trying to match an existing product profile. What tools are available to achieve this goal? What do you think about glycoengineering? Do you think it is better to try and control the glycosylation or engineer it?

The term glycoengineering has been changing over time.  Glycoengineering was traditionally defined as any tool used to modify or change the glycosylation patterns. There are 4 basic levers to pull to control the addition of glycans.  They are (1) the substrate – namely the sugar nucleotide mix and quantity, (2) the enzymes – glycotransferases and […]» Read More

What impact does glycosylation have on the protein?

Glycosylation is a critical quality to consider for proteins.  The amount and type of glycosylation has dramatic effect on the physical nature of the protein and its biological function.  In addition to amino acid sequence, glycosylation can affect the folding of proteins and their ability to aggregate with each other.  The different folding and aggregate […]» Read More

Do you find that glycosylation is an issue mainly in biosimilars? It seems everything I read on this talks about trying to match the profile of an innovator product?

Conceptually this questions addresses the issue of targeting a particular glycan profile.  It is true that there is a target profile for both innovator biologics and biosimilars.  For the biosimilars, the target is clear – it is the glycan profile of the innovator. For the innovator, in the case of antibodies, there may be a […]» Read More

What do you think are the media ingredients that influence glycosylation?

This is a complex question.  There are direct effects and indirect effects.  The direct effects are mostly driven by sugar mix, specifically what kind of sugar (ex. glucose or galactose) and what form it is in.  Additionally, there are many indirect effects that control the environment.  The amount of a certain amino acid such as […]» Read More

In your description, you mention that Cell-Ess improves the consistency of glycosylation patterns. Have you seen this replicated and what kind of media are you adding your supplement to? I am just wondering about the consistency of the media and whether that would affect your results. Thanks

You ask an interesting question.  We did see the same effect of increased consistency in two different media bases when adding Cell-Ess.  Interestingly as you imply there were two different glycosylation patterns because the constituents in the base media were different but the increase in consistency of the glycoform patterns within each group remained the […]» Read More

What factors influence glycosylation?

Glycosylation is affected by several factors, including: movement of the protein through the ER and Golgi apparatus, environmental factors, and sugar availability.  The movement of protein through ER and Golgi apparatus can be affected by cholesterol and other free fatty acid availability that impact health of ER and Golgi membranes.  Environmental factors include cell culture […]» Read More