Controlling Disulfide Bond Formation in Antibody Solutions
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Solution Overview
Problem
Current biomanufacturing processes struggle to effectively control the formation of disulfide bonds between polypeptides in recombinant proteins, particularly in antibodies like IgG4, leading to the production of half-antibodies which can affect protein stability and functionality.
Innovation Solution
A methodology involving the use of conditioned solutions with specific redox reagents, pH, and temperature to control the formation of disulfide bonds between polypeptides, allowing for the adjustment of the number of disulfide bonds in proteins during bioprocesses such as batch, semi-continuous, or continuous bioprocesses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional biomanufacturing processes are used for producing IgG4 antibodies, then production efficiency is maintained, but the proportion of half-antibodies increases significantly (at least 35%)
Solution Approach 1:
The patent applies parameter changes by systematically varying pH, temperature, ionic strength, and redox potential in the incubation buffer to optimize disulfide bond formation. Specific embodiments use pH 7.0-7.5, temperature 25-37°C, and specific redox buffers to convert half-antibodies to full antibodies while maintaining production efficiency
Solution Approach 2:
The patent uses intermediary substances including redox buffers (such as glutathione, TCEP, or DTT), chaotropic agents (such as guanidine hydrochloride or urea), and specific ions (such as zinc or calcium) to mediate the conversion of half-antibodies to full antibodies during the incubation process
2Reliability
If IgG4 antibodies are produced without controlled disulfide bond formation, then manufacturing process is simpler, but protein stability and functionality are compromised
Solution Approach 1:
The patent applies preliminary action by adding half-antibody conversion steps during the manufacturing process, such as incubating with reduction agents followed by oxidation, or using engineered cell lines that naturally promote correct disulfide bond formation, thereby ensuring protein stability before final product formulation
Solution Approach 2:
The patent employs self-service mechanisms through engineered expression systems that naturally facilitate correct disulfide bond formation, such as using specific host cells with optimized protein folding environments, chaperone proteins, or post-translational modification systems that automatically correct half-antibody formation
3Adaptability or versatility
If half-antibodies are formed in IgG4 production, then dynamic Fab arm exchange capability is enhanced, but aberrant protein formation occurs
Solution Approach 1:
The patent converts the harmful effect of half-antibody formation into a beneficial feature by deliberately controlling the process to maintain specific proportions of half-antibodies for therapeutic purposes, such as creating bispecific antibodies or enhancing antigen binding versatility, while using purification and formulation steps to eliminate aberrant proteins
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 method enables the precise control of disulfide bond formation, reducing the proportion of half-antibodies, enhancing protein stability and consistency in therapeutic antibody production.
Implementation Method 1
the conditioned solution comprises one or more predetermined solution parameters... the incubation of the polypeptides with the conditioned solution controls the formation of disulfide bonds between the polypeptides of the protein
Data Source
AI summary
Disclosed herein are methods that have been developed to control the formation of disulfide bonds between polypeptides of a multimeric protein produced by a bioprocess. Also disclosed are protein solution parameters that allow for controlling the formation of disulfide bonds. In one example, the methods disclosed herein can be used to control the proportion of half antibody molecules in an antibody solution.