High-Concentration Liquid Medium Preparation at Reduced Osmolarity
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Solution Overview
Problem
Conventional methods for preparing high-concentration cell culture media result in increased osmotic pressure due to the use of large amounts of acid and base, which can negatively impact protein productivity in animal cell cultures.
Innovation Solution
A method involving hydrating first and second medium components at specific temperatures, followed by cooling and mixing, to prepare a high-concentration liquid medium with reduced osmotic pressure, using a solvent at 35 °C or more and adjusting pH to between 6.5 and 7.5.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If high pH is applied to dissolve medium components, then components that do not readily dissolve can be dissolved, but large amounts of acid and base must be added which causes substantial increase in osmotic pressure
Solution Approach 1:
The patent applies parameter changes by heating the solvent to 35°C or more before mixing with medium components, and adjusting pH to 6.5-7.5 instead of using high pH (8-9). This changes the physical and chemical parameters of the system to achieve dissolution without requiring excessive base and subsequent acid neutralization, thereby reducing osmotic pressure while maintaining ease of manufacture
Solution Approach 2:
The patent replaces the conventional mechanical/chemical approach of using high pH to force dissolution with a thermal approach (heating to 35°C or more) combined with controlled pH adjustment (6.5-7.5). This substitution eliminates the need for large amounts of base and acid, reducing the osmotic pressure increase while still achieving complete dissolution of medium components
2Productivity
If high density cell seeding is applied to increase protein yield, then productivity increases, but nutrients must be supplied in quantities proportional to increased cell density which increases osmotic pressure
Solution Approach 1:
The patent changes the physical parameters of the medium by heating the solvent to 35°C or more and adjusting pH to 6.5-7.5, which allows high-concentration medium preparation with reduced osmotic pressure. This enables high-density cell seeding (for increased productivity) without the osmotic pressure increase that would normally result from adding large quantities of nutrients
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method effectively reduces osmotic pressure, maintaining cell viability and protein productivity by minimizing the use of acid and base, resulting in a medium with decreased osmolarity.
Implementation Method 1
mixing a first medium component with a solvent at a temperature of 35 °C or more to hydrate the first medium component
Implementation Method 2
cooling a solution containing the hydrated first medium component
Implementation Method 3
mixing a second medium component with the solution to hydrate the second medium component
Data Source
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AI summary
The present disclosure relates to a method of preparing a high-concentration liquid medium having reduced osmolarity, and a method of producing a target protein using the medium. A high-concentration liquid medium, when prepared according to one aspect, may have effectively reduced osmolarity. Therefore, the medium, when applied to a process for producing a target protein, may increase the production/expression level of the target protein, increase cell count, increase cell viability, and increase specific productivity of cells, thereby significantly increasing the production yield of the target protein from cells.