Rapid High-Throughput Feed Optimization - With high-throughput feed optimization, researchers can achieve in a single experiment what it would take months to do using shake flasks. - BioPharm

ADVERTISEMENT

Rapid High-Throughput Feed Optimization
With high-throughput feed optimization, researchers can achieve in a single experiment what it would take months to do using shake flasks.


BioPharm International


RESULTS

Component Screening


Table 1. Combinations from the 16-component central composite design
Dilution series of selected feed components were performed to check for optimal concentration ranges and toxicity levels. These experiments were performed in a shaker plate system. A serial dilution was performed from 2x through 0x. The plates were incubated at 37C, 5% CO2, and were run for 7 to 9 days. Cells were sampled on days 3, 5, 7, and 9 for cell viability assays. For one set of experiments, it took one day to set up the experiment; after incubating the plates for 7 to 9 days, significant components were selected based on cell viability and protein productivity assays. A schematic of the media development strategy is shown in Figure 1.

Sixteen-Factor Central Composite Design


Figure 2. Cell viabilities of the control, run 258, and run 179 in triplicates are shown from day 0 to day 9. The controls maintained good viability for 5 days and samples 258 and 179 maintained good viability for 7 to 8 days.
Approximately 16 components were selected for feed optimization based on the cell growth curves from feed formulation experiments. A 16-component central composite design was set up in triplicates in a shaker plate system. Cells were seeded at a density of 1.85 x 106/mL, and the plates were incubated at 37C, 5% CO2, and were run for 7 to 9 days. Cells were sampled on days 3, 5, 7, and 9 for high-throughput cell viability and protein productivity assays. Two of the top combinations from the shaker plate system were selected for shake flask studies based on protein concentration. The factorial design of some of the combinations is shown in Table 1.

Shake Flask Studies


Figure 3. Viable cell counts of the control, run 258, and run 179 in triplicates are shown from day 0 to day 9. The maximum viable cell count for the controls was on day 5 and for runs 179 and 258 was on day 6 and day 7, respectively.
Shake flask studies were performed with combinations 179 and 258, as shown in Table 1, to confirm the results from the shaker plate system. These studies were performed in triplicates along with the controls. The feed components were added according to the combinations, and the controls were added with the same amount of media but no additional components. This process was followed to ensure the same volumes in all the experimental shake flasks. The initial seeding density after adding the components was 1.72 x 106/mL. Cells were sampled on each day up to 9 days. Cell viability and cell counts are shown in Figures 2 and 3. The controls reached a maximum viable cell density of 4 x 106/mL on day 5 and dropped below 2 x 106/mL with 45% viability on day 6.


blog comments powered by Disqus

ADVERTISEMENT

ADVERTISEMENT

Pfizer to Acquire Vaccines from Baxter
July 30, 2014
GSK Submits EU Regulatory Filing for Malaria Vaccine Candidate
July 29, 2014
Bristol-Myers Squibb and Ono Pharmaceutical Collaborate on Immunotherapies
July 28, 2014
FDA Accepts First Biosimilar Filing
July 24, 2014
Compounding Pharmacy Issues Recall, But Challenges FDA Decision
July 22, 2014
Author Guidelines
Source: BioPharm International,
Click here