Chinese hamster ovary (CHO) is the cell line of choice in many biotherapeutic production workflows. Yet, in the early stages of drug development, CHO lines require high-performing, chemically defined media optimized for their specific needs.
While media optimization can be both expensive and time consuming, choosing a preoptimized CHO feedstock can improve recombinant protein production and help companies stay competitive in the drug development field.
This application note highlights the effectiveness of Gibco™ Efficient-Pro™ media in maintaining CHO health and productivity for biotherapeutics applications.
Download this application note to discover:
- A range of media options for various CHO cell lines
- Feed system designs informed by the latest multiomics data
- Preoptimized approaches that work like custom solutions
Chinese hamster ovary (CHO) cells are the primary cell line for production of biotherapeutic proteins, accounting for up to 70% of recombinant proteins [1]. These cells are the workhorse for bioproduction because they allow for human-like posttranslational modifications and are not susceptible to infection by human viruses [2]. Year after year, biopharmaceutical manufacturers have successfully driven their CHO cells to deliver increased productivity. Despite these advances, the industry continues to strive toward maximizing productivity by streamlining manufacturing processes to reduce costs. To maximize productivity and reduce development time and costs, CHO workflows in the early stages of development require a higher-performing chemically defined (CD) platform with basal media and feeds optimized for specific CHO cell lines. To help provide solutions for the bioproduction industry, the Gibco™ Efficient-Pro™ Medium and Gibco™ Efficient-Pro™ Feeds 1 and 2 were developed using a traditional and multiomics modeling approach to improve recombinant protein production in CHO cells. The CD Efficient-Pro Medium is supplemented with either of the corresponding single-part feeds designed for optimal performance with specific CHO cell lines. Efficient-Pro Feed 1 is designed primarily for CHO-K1 cells and EfficientPro Feed 2 for CHO-S and DG44 cells. The performance of Efficient-Pro Medium and Feeds was evaluated using CHO-K1, CHO-S, and DG44 cells and compared to another supplier’s commercially available basal medium and 2-part feed. The cell lines were evaluated in a 14-day fed-batch IgG productivity study with cell growth, viability, titer, and specific productivity assessed at multiple time points. In addition, critical metabolites that are known to impact cell health were evaluated. Table 1. Feed supplementation strategy. CHO-K1 CHO-S DG44 Efficient-Pro Feed 3% Feed 1 2.5% Feed 2 1.5% Feed 2 Other supplier’s feeds 3% Feed A and 0.3% Feed B 3% Feed A and 0.3% Feed B 3% Feed A and 0.3% Feed B Materials and methods Cell culture In-house CHO-K1, CHO-S, and DG44 cells expressing IgG were recovered in banking medium, then adapted for a minimum of 3 passages in each test basal medium in shake flasks. Cultures were expanded and set up in triplicate for each condition in an Ambr™15 bioreactor (Sartorius), with a seeding density of 0.3 x 106 viable cells/mL. Efficient-Pro Medium (Cat. No. A5322201) and another supplier’s basal medium were supplemented with 6 mM L-glutamine and 1:100 Gibco™ Anti-Clumping Agent (Cat. No. 0010057). Culture conditions were set at pH 7.05, 50% DO, 37°C, and 1,200 rpm for all cell lines, except CHO-K1, which was cultured at 1,000 rpm on days 1–9 and 1,200 rpm on days 10–14. The cultures were supplemented daily on days 3–13 with either Efficient-Pro Feed 1 (Cat. No. A5208801), Efficient-Pro Feed 2 (Cat. No. A5221404), or the other supplier’s two feeds, as outlined in Table 1. Glucose was fed to 6 g/L when the concentration dropped below 3.5 g/L. Cell counts and viability were evaluated using a Vi-CELL™ XR Analyzer (Beckman Coulter). Note: Cultures were supplemented daily on days 3–13. The recommended daily concentration of Efficient-Pro Feed ranges from 1.5% to 3% and can be optimized depending on the specific nutritional requirements of each cell line. The other supplier’s feed concentrations were based on the manufacturer’s recommendations.0 10 20 30 40 50 60 70 80 90 100 0 10 20 30 40 3 5 7 10 12 14 Average viability (%) Average VCD (x 106 cells/mL) Days CHO-S growth and viability Other supplier’s medium + feeds (VCD) Other supplier’s medium + feeds (viability) E‹cient-Pro Medium + Feed 2 (viability) E‹cient-Pro Medium + Feed 2 (VCD) Relative average qP (%) Other supplier’s medium + feeds (qP) Other supplier’s medium + feeds (titer) 200 150 100 50 0 Relative average titer (%) 200 150 100 50 0 3 5 7 10 12 14 Days E‹cient-Pro Medium + Feed 2 (titer) E‹cient-Pro Medium + Feed 2 (qP) CHO-S IgG productivity 0 10 20 30 40 50 60 70 80 90 100 0 10 20 30 40 50 60 3 5 7 10 12 14 Average viability (%) Average VCD (x 106 cells/mL) Days CHO-K1 growth and viability Other supplier’s medium + feeds (VCD) Other supplier’s medium + feeds (viability) E‹cient-Pro Medium + Feed 1 (VCD) E‹cient-Pro Medium + Feed 1 (viability) 0 20 40 60 80 100 120 140 160 180 Relative average qP (%) Other supplier’s medium + feeds (qP) Other supplier’s medium + feeds (titer) 3 5 7 10 12 14 Days E‹cient-Pro Medium + Feed 1 (qP) E‹cient-Pro Medium + Feed 1 (titer) 0 20 40 60 80 100 120 140 160 Relative average titer (%) CHO-K1 IgG productivity Titer, metabolites, and specific productivity Antibody titers, lactate, and ammonia metabolites were assessed using a Cedex™ BioHT Analyzer (Roche). Specific productivity (qP) was calculated as IgG produced, in pg/cell/day, based on Equation 1, where [IgGt0 ] and [IgGt1 ] represent the IgG product concentration on t0 or t1, and VCDt0 and VCDt1 represent the total viable cell number on t0 or t1, with t representing time in days. Equation 1 qP = ([IgGt1 ] – [IgGt0 ]) / ((VCDt1 – VCDt0 ) / ln(VCDt1 /VCDt0 ) x (t1 – t0)) Figure 1. CHO-K1 growth and productivity. (A) CHO-K1 cells had comparable average viability when grown with Efficient-Pro Medium and Feed 1 or with another supplier’s medium and feeds, but lower average peak VCD with the Efficient-Pro medium and feed. (B) By day 14, relative average titers were comparable in both media, but relative average qP was higher, up to 170%, with the Efficient-Pro medium and feed. Percent relative average titer was based on the other supplier’s day 14 average peak titer. Percent relative average qP was based on the other supplier’s combined average qP for days 7, 10, 12, and 14. The other supplier’s system showed high variability in titer at days 12 and 14 (error bars). Figure 2. CHO-S growth and productivity. (A) CHO-S cells had comparable average cell viability when grown with Efficient-Pro Medium and Feed 2 or with another supplier’s medium and feeds, and higher average peak VCD with the Efficient-Pro medium and feed. (B) The Efficient-Pro system produced a 144% relative average titer by day 14, and up to 180% relative average qP. Percent relative average titer was based on the other supplier’s day 14 average peak titer. Percent relative average qP was based on the other supplier’s combined average qP for days 7, 10, 12, and 14.
