CD3-Binding Polypeptide Composition with Controlled Multimer Content
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Solution Overview
Problem
Recombinantly produced polypeptides that bind to the human CD3 antigen can activate cytotoxic T cells in a clonally independent manner, leading to unintended biological activities and reduced efficacy due to the formation of multimeric forms which exhibit diverse biological activities, complicating therapeutic regimens.
Innovation Solution
A composition comprising a polypeptide with two antigen binding sites, where one site specifically binds the human CD3 antigen and the other can target any antigen of interest, with a controlled minimal amount of multimeric form (≤5% of total weight) to ensure homogeneous biological activity, primarily activating cytotoxic T cells in the presence of target cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If polypeptides are produced recombinantly with multiple antigen binding sites, then therapeutic potential and ability to activate cytotoxic T cells is improved, but formation of multimeric forms occurs which causes diverse biological activities and reduces therapeutic efficacy
Solution Approach 1:
The patent applies parameter changes by controlling the oxidation state and disulfide bond formation during polypeptide production. By adjusting redox conditions and using specific buffering systems, the patent maintains polypeptides in a reduced state to prevent unwanted multimeric form formation, thereby eliminating diverse biological activities while preserving therapeutic efficacy
Solution Approach 2:
The patent applies local quality by creating different structural states within the polypeptide population - specifically controlling which cysteine residues form disulfide bonds. By using reducing agents during production, the patent ensures that only necessary intramolecular disulfide bonds form while preventing intermolecular bonding, thus eliminating multimeric forms and their associated harmful biological activities
2Productivity
If polypeptides are produced in multimeric forms, then production yield may be improved, but biological activity becomes heterogeneous which complicates therapeutic regimens
Solution Approach 1:
The patent applies parameter changes by controlling pH, redox potential, and the presence of reducing agents during production and purification. These parameter changes prevent multimeric form formation while maintaining high production yields, ensuring that the biological activity remains homogeneous throughout the therapeutic regimen
Solution Approach 2:
The patent applies preliminary action by incorporating reducing agents and controlling oxidation conditions during the production phase itself, rather than attempting to separate multimeric forms afterward. This preliminary prevention of multimeric formation ensures both high productivity and homogeneous biological activity from the outset
Data Source
AI summary
The present invention relates to compositions comprising polypeptides, especially polypeptides capable of specifically binding predetermined antigens. The polypeptide in the composition comprises at least two antigen binding sites. These at least two antigen binding sites are located on a single polypeptide chain. One of the at least two antigen binding sites specifically binds the human CD3 antigen. The polypeptide may exist in both monomeric form and multimeric form. The multimeric form of the polypeptide constitutes no more than 5% of the total weight of the combined monomeric and multimeric forms of said polypeptide.