Continuous Chromatography for Therapeutic Protein Heterogeneity
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Solution Overview
Problem
Current methods for producing therapeutic proteins face challenges in reducing heterogeneity, particularly before purification, which complicates the isolation and purification processes and increases costs due to the need for enzyme optimization and high enzyme costs in batch processes.
Innovation Solution
A continuous bio-manufacturing process utilizing a multi-column chromatography system that includes columns with carboxypeptidase B immobilized on sepharose to reduce heterogeneity in therapeutic proteins, capturing, purifying, and polishing the proteins in a single system, allowing for reduced basic isoforms and viral inactivation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If carboxypeptidase enzyme is added directly to bioreactor or after chromatographic step to reduce heterogeneity, then heterogeneity of therapeutic protein is reduced, but enzyme cost increases and process optimization complexity increases
Solution Approach 1:
The carboxypeptidase enzyme is immobilized on chromatography resin before the chromatographic step, allowing heterogeneity reduction to occur during the capture step itself rather than requiring separate optimization steps afterward. The enzyme is pre-positioned on the resin beads, creating a ready-to-use system that automatically performs both capture and heterogeneity reduction in one operation.
Solution Approach 2:
The chromatography resin acts as an intermediary carrier that holds the carboxypeptidase enzyme in a fixed position. This intermediary structure allows the enzyme to remain stationary while processing flows through it, enabling continuous operation without requiring enzyme addition or removal steps. The resin mediates between the enzyme and the therapeutic protein stream.
2Manufacturing precision
If carboxypeptidase enzyme is added directly to bioreactor or after chromatographic step to reduce heterogeneity, then heterogeneity of therapeutic protein is reduced, but overall process cost increases due to high enzyme cost
Solution Approach 1:
The chromatography resin with immobilized enzyme is designed as a disposable or easily replaceable component. Rather than investing in expensive, complex enzyme recovery and regeneration systems, the patent utilizes the relatively low-cost resin matrix that can be discarded or regenerated through simple washing steps, making the overall process more economical despite the enzyme cost.
Solution Approach 2:
The patent combines the enzyme immobilization with the chromatography resin in a single integrated step. The enzyme is covalently or non-covalently attached to the resin during resin preparation, creating a unified reagent that performs both capture and heterogeneity reduction functions simultaneously, eliminating the need for separate enzyme addition and recovery operations.
3Manufacturing precision
If batch process with enzyme addition is used to reduce heterogeneity, then heterogeneity is reduced, but processing time increases and productivity decreases
Solution Approach 1:
The immobilized enzyme on chromatography resin enables continuous processing where therapeutic protein flows continuously through the column, and heterogeneity reduction occurs continuously during the flow-through process. There are no batch incubation periods or separate enzyme addition steps, maintaining continuous productive action throughout the operation.
Solution Approach 2:
The enzyme is pre-immobilized on the resin in advance, so when the therapeutic protein flows through the column, heterogeneity reduction begins immediately without requiring preliminary incubation or activation steps. The system is pre-configured to perform the reduction function as part of the normal flow-through process.
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
This process achieves continuous production of therapeutic proteins with reduced heterogeneity, high yield, and lower enzyme usage, resulting in a more flexible and economic manufacturing process with streamlined operations and reduced capital costs.
Implementation Method 1
Carboxypeptidase B is a carboxypeptidase that preferentially acts upon basic amino acids, such as arginine and lysine
Implementation Method 2
A continuous bio-manufacturing process utilizing a multi-column chromatography system that includes columns with carboxypeptidase B immobilized on sepharose
Data Source
AI summary
The present disclosure relates to a bio-manufacturing process that enables continuous production of a therapeutic protein with reduced heterogeneity. The bio-manufacturing process disclosed herein utilizes a multi column chromatography system that performs the unit operations of reducing heterogeneity in a therapeutic protein, capturing the therapeutic protein and inactivating viruses, purifying the therapeutic protein, and polishing the purified therapeutic protein. The process disclosed herein can reduce heterogeneity of a therapeutic protein by reducing a proportion of basic isoforms of the therapeutic protein.