MBP20571a

One Perfusion Medium for Every Stage of Your Workflow

Gibco High-Intensity Perfusion (HIP) CHO Medium is an innovative, chemically defined, animal-origin free (AOF) medium. This easy-to-use perfusion medium provides high titers in numerous CHO cells from beginning to the end of the process regardless of the type of perfusion platform, all while using less medium. With the largest global manufacturing network providing assurance of supply, lot-to-lot consistency, and global support, you can focus on getting your product to market.

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  • Tested in CHO-S, CHO-GS, CHO-K1, and CHO-DG44
  • Tested in intensified fed-batch, concentrated fed batch, N-1, and continuous perfusion
  • Advanced Granulation Technology (AGT) dry media format enables a simple and scalable reconstitution process–just add water

N-1 Intensified perfusion is typically used for ramping up and concentrating cell density.  This enables skipping stages in the seed train while seeding a higher density in a production reactor.

N-1 perfusion with two different clones
Figure 1. N-1 perfusion applied to two different clones to accelerate cell growth rate at 1 VVD medium exchange rate.

In these processes, perfusion is used to achieve high cell densities while maintaining log growth.  This is a short duration method (usually 4–7 days) that is used to reduce the number of vessels required in a seed train, allow for higher density reactor seeding, or for generating high density seed banks.  Medium exchange rates are selected to maintain the culture in log growth; productivity is not usually a consideration.

In a concentrated fed batch, a smaller pore size filter is used to retain the end product. This type of perfusion is similar to fed-batch with a comparable risk of product quality issues.

Comparison of simple fed batch with a fed batch process
Figure 2. Side-by-side application of enhancing a legacy simple fed batch (glucose) with a fed-batch process and concentrated perfusion-based fed-batch process. The concentrated fed batch was operated at 1.2 maximum VVD.

With concentrated fed batch, filtration must be used as the retention method, as both the cells and the product are returned to the reactor throughout the run. This allows for concentration of the product in the vessel and an increased final titer. This is ideal for stable products with low productivity in batch or fed-batch processes; the concentrated final titer allows for more productive downstream batch processing. These are typically 14–20 day runs, and media exchange rates are targeted to generate enough titers for downstream processing.

Intensified fed batch has a moderate length run (16–22 days) and a higher than normal VCD (Viable Cell Density) and productivity.

Intensified fed batch is similar to concentrated fed batch except the product is removed from the reactor throughout the run, making this is a better option when working with more labile products.  Media exchange can lead to higher titer production, and the shorter residence time can provide better quality control than batch operation. Constant product removal provides the option of semi-continuous downstream purification as well, with pauses needed only between runs. The process run time is generally a bit longer than a standard fed batch run (16–25 days), and media exchange rates are targeted to maximize cost per titer, barring equipment limitations.

Continuous perfusion has a 30- to 90-day production run with the ability to fine tune culture process and product quality. This type of perfusion allows for the highest optimization of downstream function.

Continuous perfusion
Figure 3. Continuous perfusion at 1 VVD medium exchange rate.

Continuous perfusion maintains a steady state where productivity and product quality can be sustained long term with minimal variability. These processes can extend from 60 to 90 days, and an active bleed is employed to maintain a targeted viability percentage or VCD.  Ideally, this bleed is minimized to limit product loss and maximize the net yield of the process.  Similar to intensified fed batch, media exchange rates are optimized to maximize profits.  This the most technically demanding type of perfusion. It also requires a cell line with superior stability. Despite the technical challenges, continuous perfusion is attractive as it can provide an avenue to long-term continuous downstream bioprocessing.

N-1 Intensified perfusion is typically used for ramping up and concentrating cell density.  This enables skipping stages in the seed train while seeding a higher density in a production reactor.

N-1 perfusion with two different clones
Figure 1. N-1 perfusion applied to two different clones to accelerate cell growth rate at 1 VVD medium exchange rate.

In these processes, perfusion is used to achieve high cell densities while maintaining log growth.  This is a short duration method (usually 4–7 days) that is used to reduce the number of vessels required in a seed train, allow for higher density reactor seeding, or for generating high density seed banks.  Medium exchange rates are selected to maintain the culture in log growth; productivity is not usually a consideration.

In a concentrated fed batch, a smaller pore size filter is used to retain the end product. This type of perfusion is similar to fed-batch with a comparable risk of product quality issues.

Comparison of simple fed batch with a fed batch process
Figure 2. Side-by-side application of enhancing a legacy simple fed batch (glucose) with a fed-batch process and concentrated perfusion-based fed-batch process. The concentrated fed batch was operated at 1.2 maximum VVD.

With concentrated fed batch, filtration must be used as the retention method, as both the cells and the product are returned to the reactor throughout the run. This allows for concentration of the product in the vessel and an increased final titer. This is ideal for stable products with low productivity in batch or fed-batch processes; the concentrated final titer allows for more productive downstream batch processing. These are typically 14–20 day runs, and media exchange rates are targeted to generate enough titers for downstream processing.

Intensified fed batch has a moderate length run (16–22 days) and a higher than normal VCD (Viable Cell Density) and productivity.

Intensified fed batch is similar to concentrated fed batch except the product is removed from the reactor throughout the run, making this is a better option when working with more labile products.  Media exchange can lead to higher titer production, and the shorter residence time can provide better quality control than batch operation. Constant product removal provides the option of semi-continuous downstream purification as well, with pauses needed only between runs. The process run time is generally a bit longer than a standard fed batch run (16–25 days), and media exchange rates are targeted to maximize cost per titer, barring equipment limitations.

Continuous perfusion has a 30- to 90-day production run with the ability to fine tune culture process and product quality. This type of perfusion allows for the highest optimization of downstream function.

Continuous perfusion
Figure 3. Continuous perfusion at 1 VVD medium exchange rate.

Continuous perfusion maintains a steady state where productivity and product quality can be sustained long term with minimal variability. These processes can extend from 60 to 90 days, and an active bleed is employed to maintain a targeted viability percentage or VCD.  Ideally, this bleed is minimized to limit product loss and maximize the net yield of the process.  Similar to intensified fed batch, media exchange rates are optimized to maximize profits.  This the most technically demanding type of perfusion. It also requires a cell line with superior stability. Despite the technical challenges, continuous perfusion is attractive as it can provide an avenue to long-term continuous downstream bioprocessing.

Bioprocessing resources

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