Fed-Batch Bioreactor Feed Optimization for Lactate Spike Control
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Solution Overview
Problem
In fed-batch processes for producing proteins, a lactate spike occurs due to high pyruvate concentrations, leading to increased glucose consumption, reduced product expression, and lower product quality.
Innovation Solution
Reducing the pyruvate concentration in feeds to cells in a bioreactor by 65% to 100% and increasing the concentration of certain amino acids like glutamate, valine, and leucine in the feeds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high pyruvate concentration is used in feeds, then cell growth is supported, but lactate spike occurs leading to reduced product expression and lower product quality
Solution Approach 1:
The patent changes the concentration parameter of pyruvate in the feed from high to reduced levels (65-100% reduction). This parameter change prevents lactate spike while maintaining cell growth through compensatory amino acid supplementation, thereby resolving the contradiction between productivity and product quality
Solution Approach 2:
The patent introduces amino acids (glutamate, valine, leucine, threonine, aspartate, isoleucine) as intermediary substances that mediate between pyruvate reduction and cell growth maintenance. These amino acids compensate for the reduced pyruvate, allowing cell growth to continue without causing lactate spike, thus resolving the contradiction
2Productivity
If high pyruvate concentration is used in feeds, then cell growth is supported, but glucose consumption increases
Solution Approach 1:
The patent reduces pyruvate concentration parameter in feeds by 65-100%, which directly addresses the excessive glucose consumption caused by high pyruvate. This parameter change modifies the metabolic pathway to reduce unnecessary glucose uptake while maintaining cell growth through amino acid supplementation
Solution Approach 2:
Amino acids serve as intermediary substances that enable cell growth without triggering the high glucose consumption associated with high pyruvate. By introducing these intermediaries, the system achieves productivity while reducing substance loss
3Manufacturing precision
If pyruvate concentration is reduced in feeds, then lactate spike is reduced, but cell growth may be affected
Solution Approach 1:
The patent uses amino acids (glutamate, valine, leucine, threonine, aspartate, isoleucine) as intermediary substances that bridge the gap between reduced pyruvate and maintained cell growth. These intermediaries ensure that cell growth continues at productive levels even when pyruvate is reduced to prevent lactate spike
Solution Approach 2:
The patent simultaneously changes multiple parameters: reducing pyruvate concentration while increasing specific amino acid concentrations. This coordinated parameter change ensures that product quality improves through lactate reduction while productivity is maintained through amino acid-supported cell growth
Data Source
AI summary
Provided herein is a method for reducing a lactate spike and increasing titer in a fed-batch process for producing a protein of interest, comprising reducing pyruvate (e.g., sodium pyruvate) concentration in one or more feeds to cells in a bioreactor in the fed-batch process.