Immunoglobulin Sequence Modifications for Aggregation Control
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Solution Overview
Problem
The production of antibody molecules in heterologous systems often results in sub-optimal manufacturability due to aggregation issues and reduced productivity, particularly when mammalian proteins are expressed in non-natural environments, leading to instability and poor yield.
Innovation Solution
Modifying the amino acid sequence of immunoglobulin molecules by introducing substitutions in aggregation-prone segments of the VL and VH domains, as well as the CH1 constant region, to reduce aggregation propensity and enhance productivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If mammalian proteins are expressed in heterologous systems, then productivity is improved, but aggregation propensity increases and stability decreases
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid sequences at specific positions (N-terminal, C-terminal, and internal regions) to alter protein properties. These sequence modifications change the physical-chemical parameters of the immunoglobulin, reducing aggregation propensity and improving stability while maintaining antigen binding activity, thereby resolving the contradiction between productivity and stability in heterologous expression systems
2Productivity
If expression levels are increased to improve yield, then productivity increases, but aggregation propensity increases
Solution Approach 1:
The patent applies preliminary anti-action by pre-modifying the immunoglobulin amino acid sequence to reduce aggregation-prone regions before expression. By introducing substitutions at critical positions (removing aggregation-prone residues and adding stabilizing residues), the patent prevents aggregation from occurring even at high expression levels, thereby enabling high yield without the harmful aggregation effect
3Object-generated harmful factors
If amino acid substitutions are introduced to reduce aggregation, then aggregation propensity decreases, but manufacturing complexity increases
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions at specific local positions rather than throughout the entire sequence. The modifications are concentrated at critical regions (N-terminal, C-terminal, and internal aggregation-prone segments) while leaving the rest of the immunoglobulin structure unchanged, thereby reducing aggregation without unnecessarily increasing overall manufacturing complexity
Data Source
AI summary
This invention relates to the modification of the amino acid sequence of an immunoglobulin molecule at certain key positions within regions of the VH and VL FR and CDR3 domains and/or the CH1 domain which are prone to aggregation. Immunoglobulins modified as described may display improved manufacturability, for example, reduced aggregation propensity and/or increased production levels.