EX-CELL® Advanced™ CHO Fed-batch Media and Feed Platform Technical Data Sheet

Performance

Two studies were executed to evaluate EX-CELL Advanced CHO Fed-batch Platform in which the first study assessed indicative performance measures (viable cell density and titer) and the second study for resulting protein quality as demonstrated by relative glycan distribution (%).

Study 1 Design

Three distinct model CHO cell lines were selected for a comparative evaluation (CHO-S, CHO-dhfr, CHOZN® GS) of leading commercial growth and feed media to the EX-CELL Advanced CHO Fed-batch Media Platform by SAFC for viable cell densities and productivity. The initial adaptation step served as a basis for selecting the top competitive media for each model cell line to conduct the comparative studies. Only those media successfully adapted within three passages were selected for subsequent growth and productivity analysis. In all instances and regardless of cell bank medium, the cultures easily adapted to EX-CELL Advanced CHO Fed-batch Media Platform. All cultures were monitored for viable cell density and measured for antibody (IgG) titer up to day 14.

Culture Conditions

  • TPP tube cultures
    − 200rpm / 50mm orbit rotation
    − 5% CO2, 37°C, 80% RH
  • 1L bioreactors
    − 150rpm with 40% DO

Feed Strategy

  • 7.5% (v/v) initial culture volume
  • Supplement days 3, 5, 7, 9 and 11
  • 4 g/L glucose added when glucose less than 2g/L


Chart 1: Adaptation Scheme

Study 1 Results: VCD, Titer & Scalability

CHO S Cell Line

Graph 1: Viable cell density and productivity comparative at TTP tube scale


CHO dhfr Cell Line

Graph 2: Viable cell density and productivity comparative at TTP tube scale


CHOZN® GS Cell Line

Graph 3: Viable cell density and productivity comparative at TPP tube scale


CHOZN® GS Cell Line: Scale-up

Graph 4: Demonstrates the scalability in the performance of the EX-CELL Advanced media platform. Study comparative between SAFC’s small scale model (TPP tubes) and 1L bioreactor using the leading competitive media and feed products in the CHOZN cell line.

Protein Quality

Study 2 Design

EX-CELL Advanced CHO Fed-Batch System in combination with our Protein Quality Supplement, (Product No. 14701C) was evaluated for impact on protein quality. CHO GS cells were banked, thawed, and passaged 3 times.  Cultures were grown as previously described, and fed with EX-CELL Advanced Feed 1 on days 3, 5, 7, 9 and 11.  Glucose was supplemented with 4g/L whenever glucose levels fell below 2 g/L in culture. Additionally, EX-CELL Glycosylation Adjust (GAL+) was supplemented beginning on day 2 and every other day through day 10 of the 14 day fed-batch culture. Culture supernatants were harvested on Day 14 for purification and analysis. No differences were observed in Viable Cell Density, Viability, and Titer.

Directing Glycosylation in EX-CELL Advanced Fed-batch System

Conclusion

Study 1

The EX-CELL Advanced CHO Fed-batch platform demonstrated a sustained high biomass with maximum titers across all model cell lines. In each comparative, the growth media proved to be the only formulation with a trouble-free adaptation from hydrolysate containing or other chemically-defined formulations. Moreover the optimized media and feed combination by SAFC outperformed other leading commercial fed-batch products with a 2-4 fold increase in titer over all others. Scalability of the EX-CELL Advanced CHO Fed-batch platform was also demonstrated in a direct comparative of performance from SAFC's validated fed-batch culture model in TPP to 1 liter bioreactor yielding >4.5 g/L IgG titer.

Study 2

The EX-CELL Advanced CHO Fed-batch platform is responsive to simple and effective means for directing consistent product quality as demonstrated by the supplementation of the culture with EX-CELL Glycosylation Adjust (GAL+).

SAFC is proud to offer a set of contemporary media products that are easy to use and give a platform performance to help achieve maximum titers and consistent product quality during development. Both media formulations have established stability profiles in both powder and liquid formats. Stability for the one-part low volume addition includes 30d stability after rehydration at room temperature to allow for flexibility and efficiency during production. The EX-CELL Advanced CHO Fed-batch Platform sets a new benchmark in fed-batch culture media for product consistency, productivity, and reliability you demand.

Product Handling & Storage

Product handling using aseptic techniques is required to avoid contamination. Do not use if cloudy or if solution has precipitated. Other evidence of deterioration may include color change, pH shift or degradation of physical or performance characteristics.

Liquid media may be stored for up to 1 year at 2 to 8°C and protected from light. Storage of feed media after rehydration (Product No. 24367C and Product No. 24368C) at room temperature, protected from light for up to 30 days. Do not use after 30 days. Refer to product label for rehydration instructions and/or our web site,

www.sigma-aldrich.com/CHOperformance

Powder medium formats should be stored in the original container at 2 to 8°C and protected from moisture. Evidence of deterioration may include clumping, color change, and/or consistency in powder.

Intended Use

This product is intended for Further Manufacturing Use in the bio-manufacturing industry. It is not intended or approved for in-vitro diagnostic use in the human or veterinary industries.

Formulation Support

SAFC is committed to providing all necessary support for propritary products including EX-CELL Advanced CHO Fed-batch Platform. The disclosure process to support Regulatory Filing is simple and available upon request. Our team will rapidly respond and address any technical concerns with the media performance. Please contact Quality Services department for any support needed.

Related Products

SAFC offers the following product which are intended for use with EX-CELL Advance CHO Fed-batch Media Platform as well as other compatible CHO Fed-batch media platforms. EX-CELL Glycosylation Adjust (GAL+) allows developers to easily achieve desired N-linked glycosylation by increasing the galactose site occupancy on the oligosaccharide to a higher level.

  Product No. Format Size
EX-CELL Glycosylation Adjust (GAL+) 14701C-100ml Liquid 100ml
EX-CELL Glycosylation Adjust (GAL+) 14701C-1000ml Liquid 1000ml
For additional information, please visit our website at www.sigma-aldrich.com/finishfaster

Materials

     
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