|
Chinese Hamster Ovary (CHO) cells are of great interest for bioprocessing and pharmaceutical research and development. These cells are robust in culture and are able to produce a variety of recombinant glycoproteins at high levels on a large scale. However, different CHO cell clones often possess diverse nutritional requirements that are unique to each clone. As a result, medium optimization for CHO cells can be very challenging, often requiring the development of a custom medium for each particular clone.
The traditional approach to media development involves titrating each medium component individually to determine the optimal level of supplementation. This process involves extensive testing conditions and can take significant amounts of time. To address this, Sigma-Aldrich has developed two different kits (Kit 1) (Kit 2) that allow you to develop optimized media formulations for your specific CHO clones faster and easier than ever before.
 CHO Medium Optimization Kit 1
CHO Kit 1 from Sigma-Aldrich provides the following benefits:
- Six unique animal-component free media, including two chemically defined formulations, designed for maximum recombinant protein production in CHO cells
- Convenient format allows for rapid screening of multiple media to select optimal formulations
- Simple yet powerful blending experiments (using DOE methodology) let researchers quickly perform further media optimization experiments
- Detailed instructions for various methods of data analysis are included
- Don’t want to perform the data analysis yourself? Send us your normalized data and we’ll do it for you!
|
| CHO Medium Optimization Kit 1 |
| Product Code |
Product Name |
Size |
| CH0001 |
CHO Medium Optimization Kit 1 |
6 x 1L |
This novel kit provides researchers a convenient format in which they can rapidly screen multiple formulations to identify which formulations work best for their particular clones. It also provides detailed protocols utilizing Design-of-Experiment (DOE) methodology to allow researchers to take the three best performing media and perform blending experiments to further optimize media specific for their particular CHO systems.
The kit consists of six diverse animal-component free media designed to maximize cell growth and recombinant protein productivity in a wide variety of CHO cell clones. Two of these six media are chemically defined and consist of differing amounts of amino acids, vitamins, salts, trace elements, recombinant human insulin, Pluronic® F-68, and other organic compounds. The remaining four media contain plant-based protein hydrolysates in addition to the above list of compounds.
CHO Kit 1 Components
- CHO Medium, animal-component free
- CHO DHFR- Medium, animal-component free
- CHO Medium, Chemically Defined, animal-component free
- CHO Medium 4, animal-component free
- CHO Medium 5, animal-component free
- CHO Medium 6, Chemically Defined, 1L animal-component free
Figure 1:
 |
Following the initial screening of the 6 media contained in this kit, researchers can perform additional blending experiments with the three best performing media. This diagram and table represents the recommended blends for this initial set of experiments for media A, B, and C.
|
Figures 2 and 3:
 |
Cell growth and viability in a recombinant CHO clone were compared under suspension conditions in blending experiments as described in Figure 1.
Samples were taken daily to determine cell growth and recombinant protein productivity. The data demonstrates that these mixing experiments can identify several blends that give increased performance over the individual starting media. |
Figure 4:
| Normalized Mixture Data
| |
| GROWTH
| |
PRODUCTIVITY
|
| 50% A, 50%B |
= 142% |
|
67% C, 17% A, 17% B |
= 367% |
| 67% A, 17% B, 17% C |
= 141% |
|
50% A, 50% C, |
= 327% |
| 67% B, 17% C, 17% A |
= 135% |
|
50% C, 50% B, |
= 327% |
| 33% A, 33% B, 33% C |
= 129% |
|
67% B, 17% C, 17% A |
= 325% |
| 67% C, 17% A, 17% B |
= 120% |
|
100% C |
= 306% |
| 50% A, 50% C |
= 119% |
|
33% A, 33% B, 33% C |
= 300% |
| 50% C, 50% B |
= 119% |
|
100% B |
= 286% |
| 100% B |
= 155% |
|
50% A, 50% B |
= 225% |
| 100% C |
= 114% |
|
67% A, 17% B, 17% C |
= 225% |
| 100% A |
= 100% |
|
100% A |
= 100% |
The data from the set of experiments shown in Figures 2 and 3 is presented in Figure 4 and is normalized to the performance of Medium A (A=100%) and ranked in terms of performance.
|
back to top
CHO Medium Optimization Kit 2
CHO Kit 1 from Sigma-Aldrich provides the following benefits:
- Minimal medium approach lets you build your own CHO basal medium
- Six optimization supplements allow for easy customization of media or feed supplements for your particular CHO clone
- Factorial matrix design allows for rapid optimization of medium formulations in far fewer conditions than traditional methods
- Detailed instructions for various methods of data analysis are included
- Don’t want to perform the data analysis yourself? Send us your data and we’ll do it for you!
|
CHO Kit 2 brings a unique minimal media approach to optimization. Utilizing a factorial matrix design (based on DOE methodology), this novel kit greatly reduces the time and effort needed to fully optimize media or feed supplements for your specific CHO clones. Furthermore, if you’re working with multiple clones, the design of this kit lets you “build your own” basal medium, and design groups of optimization supplements that are specific for each clone.
CHO Kit 2 consists of a concentrated minimal basal medium, five concentrated basal supplements, and six concentrated optimization supplements. All optimization supplements have been shown to have significant effects on CHO cell growth and/or recombinant protein production in CHO clones.
The design of this kit allows for the testing of several supplements simultaneously at high and low levels, with all possible combinations based on factorial matrix design. This design gives results much faster than the traditional approach and allows the researcher to recognize interactions between components in far fewer test conditions than with traditional methods, without losing important information.
| CHO Medium Optimization Kit 2 |
| Product Code |
Product Name |
Size |
| C 4364 |
CHO Medium Optimization Kit 2 |
6 x 1L |
CHO Kit 2 contains sufficient concentrated solutions to prepare 20 L of optimized CHO AF PF medium and consists of the following components:
CHO Kit 2 Components
- CHO Kit 2 Base Medium Concentrate Basal Supplements
- CHO Kit 2 Iron Mix
- CHO Kit 2 Glucose
- CHO Kit 2 Salt Mix
- Sodium Chloride 5M Solution
- Insulin Solution Optimization Supplements
- CHO Kit 2 Metals
- CHO Kit 2 Amino Acids Mix
- CHO Kit 2 Vitamins Mix
- CHO Kit 2 Iron Chelator Solution
- CHO Kit 2 Hydrolysates
- Fatty Acid Supplement
Figure 5:
 |
Medium optimization for amino acids, metals, and iron solution was performed for a CHO-K1 clone. The graph depicts the cell growth curves generated in all media formulations and Base Medium.
|
Figure 6:
 |
Medium optimization for vitamins, fatty acids, and hydrolysates was performed for the same CHO-K1 clone. The graph depicts the cell growth curves generated in all media formulations and Base Medium.
|
Figure 7:
 |
The cell growth curves generated in Part 2 suggest that the center point is the optimal medium. Design Expert analysis identified no significant factors, indicating that this is the optimal medium formulation for this clone.
|
Figure 8:
 |
We performed a final growth study using this optimized formulation, two competitors media and the Base Medium in CHO Kit 2. As demonstrated in the graph, over 1.5-fold increase in peak cell density and accumulated integrated cell area was achieved in the optimized formulation compared with the other three media.
|
back to top
|
