CHO Media and Feeds for Optimized Production of Therapeutic Proteins

Section Overview

• CHO Media and Feeds for Monoclonal Antibody and Next Generation Biopharmaceuticals Upstream Bioprocessing
• Media for Cell Line Development, Cell Banking and Expansion
• Media for Fed-Batch Applications
• Feeds for Fed-Batch Applications
• Feeds to Modulate Glycan Profiles in Fed-Batch Applications
  • Cellvento^® ModiFeed Platform
• Media for Perfusion Applications
• Related Products
   

CHO Media and Feeds for Monoclonal Antibody and Next Generation Biopharmaceuticals Upstream Bioprocessing

Successful production of monoclonal antibodies (mAbs) and next generation therapeutics relies on robust upstream processes that maximize productivity and ensure the right critical quality attributes without being unnecessarily complex. Key factors in achieving these goals can include selection of an optimized combination of cell culture media and feeds, use of an appropriate feeding strategy, and stringent control of culture conditions.

Depending on whether the upstream process is run in batch, fed-batch, or perfusion mode, media and feed requirements can be quite different:

• Batch mode: Cells are grown in a defined volume of medium, without media exchange or replenishment of nutrients through feeds. While the process is typically easy to set-up and to operate, productivity is limited due to the build-up of toxic by-products and depletion of nutrients.
• Fed-batch mode: Cells are initially grown in a relatively lean production medium followed by regular feed supplementation to achieve the desired product yield. Compared to batch processes, cell culture growth is increased andviability is prolonged, resulting in increased productivity.
• Perfusion mode: Medium is continuously exchanged which results in optimal nutrient conditions and delivers consistent product quality due to continuous harvest. With this intensified approach, cell culture length can be significantly prolonged compared to both batch and fed-batch modes. Perfusion operations can also be implemented in an intensified seed-train for accelerated expansion and higher cell densities. These intensified upstream processes maximize productivity.
  

Cellvento^® and EX-CELL^® CHO cell culture media and feeds are designed for high performance production of mAbs and other biopharmaceuticals in fed-batch and perfusion modes. The media and feeds are produced using highly controlled raw materials, are chemically defined, and contain no animal-derived components. Compacted media are easy to dissolve and handle with less dust, which increases operator safety while high concentration Cellvento^® feeds reduce the volume added to the bioreactor. Emprove^® dossiers are available for the media and feeds to support regulatory requirements.

The following table provides an overview of CHO media and feeds, each of which has been specifically developed for optimized performance in the respective application.

 

Table 1. Media and feeds suitable for CHO cell culture processes.

Cell Line Development	CHOZN® Electroporation, Selection, and Single Cell Cloning EX-CELL® CD Fusion medium Cellvento® 4CHO-C medium
Banking and Traditional or Perfused Seed Train	EX-CELL® CD Fusion medium	Cellvento® 4CHO-X medium
Production	Traditional and Intensified Fed-Batch	Dynamic and Steady-State Perfusion
Basal Media	EX-CELL® Advanced FB Cellvento® 4CHO	EX-CELL® Advanced HD Perfusion
Complete Feeds, Base Feeds	Cellvento® ModiFeed Prime Cellvento® 4Feed EX-CELL® Advanced Feed
Complete Feeds, Glycosylation Modulating Feeds	Cellvento® ModiFeed Gal+ Cellvento® ModiFeed Gal- Cellvento® ModiFeed Sial+

Media for Cell Line Development, Cell Banking and Expansion

EX-CELL^® CD Fusion and Cellvento^® 4CHO-X support CHO cell line expansion from vial to production.

• Cellvento^® 4CHO-C medium is a specifically developed formulation to support single cell cloning.
• EX-CELL^® CD Fusion medium is a complete medium for batch applications to support healthy cell growth in expansion. In cell line development it is combined with Cellvento^® 4CHO-C to support single cell cloning. Suitable for cell banking.
• Cellvento^® 4CHO-X expansion medium is designed to support cell expansion in batch applications and can be used in perfused seed train as it supports high cell densities. Suitable for cell banking. For more information see: Seed Train Intensification Using High Cell Density Cryopreservation
   

Media for Fed-Batch Applications

EX-CELL^® Advanced and Cellvento^® 4CHO fed-batch production media are basal formulations that support growth and productivity in CHOZN^® and multiple other CHO cell lines (e.g. CHO-K1, CHO-DG44, CHO-S) in combination with the platform feeds. Production formulations should be evaluated with the specific CHO cell line to be used in order to determine the best fit and select the appropriate feed.

• EX-CELL^® Advanced CHO Fed-batch medium is designed for use with a broad range of CHO cell lines. It is formulated with components that influence critical process parameters and product attributes such as growth, productivity, glycosylation or charge variance,
• Cellvento^® 4CHO COMP medium is a lean production medium designed to work optimally with highly concentrated feeds, increasing process efficiency.
   

Feeds for Fed-Batch Applications

Cellvento^® compacted and highly concentrated feeds extend fed-batch culture duration, improve performance, and increase titer by replenishing critical nutrients and reducing feed volume addition.

• Cellvento^® ModiFeed Prime feed is recommended for combination with Cellvento^® 4CHO and EX‑CELL^® Advanced CHO fed-batch production media. The feed maximizes the performance of suspension CHO cell lines.
  
  With ever evolving approaches to fed-batch bioprocessing, Cellvento^® ModiFeed Prime feed supports intensified processes through modification of the standard feeding schedule, generating more product in less time.
  
  Cellvento^® ModiFeed Prime feed is part of the Cellvento^® ModiFeed platform supporting a tailored feeding strategy, complemented by feeds targeting specific glycan profiles (more information in the section Feeds to Modulate Glycan Profiles in Fed-Batch Applications).
  
  
• Cellvento^® 4Feed contains cysteine and tyrosine derivatives that release free cysteine and free tyrosine slowly throughout the culture. This avoids tyrosine depletion which can lead to sequence variants and results in free cysteine release, while maintaining a reduced redox environment. Cellvento^® 4Feed is protein free.
  
  
• EX-CELL^® Advanced Feed is formulated to be combined with EX-CELL^® Advanced Fed-batch medium to replenish nutrients over the fed-batch duration. The liquid feed can be combined with Cellvento^® 4Feed to further boost CHO cell culture performance. For more information see: One -step Feed Optimization for Upstream Cell Culture Processes
   

Feeds to Modulate Glycan Profiles in Fed-Batch Applications

The Cellvento^® ModiFeed platform includes different feed formulations that can be added to CHO cell cultures to predictably and reliably modulate glycan profiles to maintain or improve protein quality. Glycan modulation feeds are compatible with CHO media formulations and can be administered as single, complete feeds or used in combination with Cellvento^® ModiFeed Prime feed to further refine glycosylation profiles. Cellvento^® feed formulations do not include glucose, which enables greater control of this key nutrient during fed-batch processes.

Table 2 outlines the available Cellvento^® ModiFeed formulations and their applications. These media support optimal cell growth and high productivity of suspension CHO cell lines such as CHO-K1, CHO-DG44, CHO-S and CHO-GS, including cell lines derived using the CHOZN^® platform.

Table 3 summarizes the features and advantages of the feeds.

Table 2. Cellvento^® ModiFeed formulations, how they are used, and the recommended companion fed-batch media.

Cellvento® ModiFeed Formulation	Application
Cellvento® ModiFeed Prime COMP feed	Single higlyh concentrated feed to improve performance; can be combined with Cellvento® ModiFeed Gal+, Gal- or Sial+ feed to tailor a desired glycosylation profile.
Cellvento® ModiFeed Gal+ COMP feed	Maximizes terminal galactosylation; higher galactosylation levels enhance complement-dependent cytotoxicity (CDC) of galactosylated IgG
Cellvento® ModiFeed Gal- COMP feed	Minimizes galactosylation; lower galactosylation levels  support biosimilar development
Cellvento® ModiFeed Sial+ COMP feed	Maximizes sialylation level; sialylation prolongs serum half-life of therapeutic proteins and enhances anti-inflammatory activity
Recommended Fed-batch Media	Cell Types
Cellvento® 4CHO COMP	All CHO cell lines, specifically CHO-K1, CHOZN®
EX-CELL® Advanced CHO Fed-batch Medium	All CHO cell lines, specifically CHOZN®

Table 3. Summary of Cellvento^® ModiFeed platform features and advantages.

Cellvento® ModiFeed Platform Features	Advantages
Chemically defined, non-animal origin (NAO) components	Minimizes risk of potential viral contamination and ensures batch-to-batch consistency
Compatible for a tailored feeding strategy	Allows a high degree of customization and flexibility in glycosylation levels
Highly concentrated formulation	Minimizes total feed volume (15–25%), resulting in maximized bioreactor working volume, decreased product dilution and increased yield
Glucose-free formulation	Allows full control of the glucose feeding strategy
Provided in compacted dry powder format	Reduces the complexity of hydration, storage and is convenient to use
Neutral pH; requires no pH adjustment	Eliminates use of acids or bases in hydration and the influence of high or low pH feeds on the fed-batch process
Hydrated medium can be stored protected from light at 2–8°C for up to 90 days or at room temperature for up to 30 days	Increases storage and use options while reducing contamination risk by limiting bioreactor connections when stored at room temperature

Media for Perfusion Applications

Cellvento^® 4CHO-X and EX-CELL Advanced^® HD Perfusion media are specifically designed for use in perfused seed train (N-1) applications and perfusion production processes.

• Cellvento^® 4CHO-X COMP Expansion medium has a high nutrient concentration and supports low cell specific perfusion rates (CSPRs) at high cell densities in N-1 perfusion applications.
• EX-CELL^® Advanced HD Perfusion medium supports growth and productivity across a diverse set of CHO cells and is designed for long term growth in dynamic and steady-state perfusion.

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