Gibco Peptones

Sources of bioprocessing variation: raw material contribution

Understanding and controlling variability is essential to having a robust manufacturing process for our customers. In this example, we dispel the common myth that peptones are the only cause of variability in a media formulation.

Figure 1. Panel A depicts the manganese levels in 23 different lots of one our most popular peptones that are used in our customers base media formulations. While there is some variation because it is a natural substance, it is within a narrow range due to our very strict process controls. Panel B illustrates a test for variation in manganese levels of a vitamin being added to the same base media and clearly depicts a much wider range of variability.

It is important to understand that most media are formulated with a variety of raw materials and trace metal contamination can be introduced via any one of them. All raw materials have variability and “chemically defined” does not mean chemically pure.

If you are going to add one key supplement to your media formulation to achieve performance, why not leverage the most nutrient rich, process-controlled supplements you can possibly get?

Leveraging peptones to achieve an acceptable glycan profile

While increasing and maintaining protein titer continues to be a key bioprocessing goal, ensuring acceptable protein quality has become an equally, if not greater, critical process specification. This is especially relevant to the ever-increasing number of biosimilar therapeutics being developed where it is absolutely essential that characteristics such as glycosylation and charge variant profiles match those of originator molecules.

Figure 2. This figure illustrates the overall glycan composition where the goal was to shift the profile. The dark blue bar reflects the chemically defined (CD) control medium. The light blue bar shows the peptone supplementing the control media and the gray and yellow show the CD media supplement.

Whether adding the peptone or either CD supplement, the control media yielded similar shifts in the profile.

A history of pioneering the future in culture media and supplements

By integrating with Thermo Fisher Scientific, we combine the trusted Difco and Gibco premier brands to provide comprehensive supplements and media solutions in one portfolio–delivering even greater value to our customers. Our long and proven history of developing peptones specifically for the bioproduction industry means we have spent many years determining the key nutrient drivers associated with cellular metabolism and fermentation. Let us show you the depth of our analytical capabilities and support you in identifying the key performance drivers in your process.

Figure 3. A history of pioneering the future.

Helping ensure consistency in processes with peptones

Need an edge? Our experience in premium supplementation is decades long and now more than ever, as part of the Gibco media portfolio, we can help accelerate your product development timeline.

Peptones are a powerful optimization tool

Our peptones are proven supplements for enhancing bioproduction applications and continue to be widely used for the production of today’s blockbuster biotherapeutics.

Figure 5. Our team optimized media with the goal of achieving a viable cell density of 10 million cells by day 5. A peptone-containing formulation was identified after the initial studies that met the design criteria. The best performing chemically defined (CD) medium required additional optimization. After an additional nine months of work, the optimized CD medium showed improved performance, but still did not meet the production goal.

Our team demonstrated that peptone containing serum replacement supplement (SRS) can result in cell performance that is comparable to 10% serum (Figure 6).

Cells cultured in serum replacement supplement (SRS) containing animal origin-free peptones divided faster with lower doubling time and high cumulative population doublings (CPD) in comparison to 10% fetal bovine serum (FBS).

Cells adapted quickly to SRS formulation from serum and no senescence of cells was observed even after 45 consecutive passages.

While there is no “drop in” replacement for serum, peptones represent one of the most nutrient rich alternatives and we can work with you to help determine what will work best in your application. And we have an entire line of animal origin-free peptones at our disposal that will likely significantly reduce your costs.

Why completely reformulate when we can offer balanced nutrition to meet the specific requirements of most cell types. And who knows? Your performance may improve!

• Select a good base medium—more enriched base media yield better results than deficient media
• Select a wide variety of peptones to evaluate—consider different substrates and multiple products from the same substrate
• Create a thorough experimental design—test multiple concentrations and individual and blended conditions
• Characterize the culture throughout the process using analytical techniques—establish baseline data using spent media in multiple runs with multiple lots of peptone
• Evaluate performance on proliferation, production, and protein quality—monitoring only one attribute is not always predictive of overall performance

Figure 7. Peptones should be incorporated as early as possible in the upstream process and at any step throughout the development cycle.

With the addition of one supplement, peptones bring numerous benefits to your cell culture.  Peptones offer versatility through a balanced composition to meet specific requirements of all cell types.

Figure 9. Peptones are versatile and can be used for a broad range of cell types.

Peptone release testing focused for bioproduction needs

Our stringent traditional and specialized testing covers a range of industries. In addition to the traditional testing performed on peptones designed for use in a range of industries, such as the food industry, we also offer specialized testing for the bioproduction market.

Peptone sources and applications

Despite the growing interest in animal origin-free peptone material, peptones from animal proteins remain effective and reliable for many cell culture and bioprocessing applications. Animal origin (AO) peptones are used in reducing or eliminating serum needs in certain media formulations, and commonly promote strong cell growth and toxin titers for vaccine production. AO peptones are derived from reliable and highly regulated raw materials and they provide a well-performing, cost-effective media component or supplement, particularly in processes and applications with less rigorous animal origin requirements.  

• Tissue-based peptones are proteins from animal sources that have been hydrolyzed into amino acids and peptides. Sources of animal protein for peptones can include various tissues, like muscle or pancreas; these tissues derive from animals highly regulated for human consumption.
• Milk-based peptones are derived from bovine milk, which is sourced under the same conditions as milk produced for human consumption. Milk is a complex material, consisting of water, lactose, lipids, salts, and proteins. Casein (80%) and whey (20%) are the fundamental protein components in milk.

Historically, microbial and mammalian cell lines have been propagated in media containing animal components, including serum. In recent years, media formulators have been moving to eliminate animal-sourced components in media formulations, motivated by BSE/TSE outbreaks in the 1990s. In response, we have developed a broad offering of yeast extracts, soy peptones, and other animal origin-free media and supplements.

Peptones derived from animal origin-free sources provide unique nutritional profiles suitable for both bacterial fermentation and mammalian cell culture applications. Yeast extracts and yeastolates offer a different range of nutritional choices to enhance production. Soy peptone, one of the first successful plant-based peptones to be optimized into cell culture, is processed in several different ways to provide various nutrient mixes to meet different needs. In addition to these more common animal origin-free components, many other plant-based products have been developed, including malt, cotton, and wheat sourced peptones, which offer an additional breadth of supplements with strong and varied performance characteristics.