Stable Soluble Human TNF Alpha Antibodies
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current anti-TNFα antibodies for treating TNF-mediated disorders, such as rheumatoid arthritis, have limitations in stability, solubility, and immunogenicity, necessitating the development of more effective and stable antibody forms for continuous treatment.
Innovation Solution
Development of stable and soluble antibodies with aggregation-reducing modifications, specifically optimized light chain and heavy chain sequences for scFv antibodies and Fab fragments, which include modifications like arginine substitution at specific positions to enhance stability and reduce immunogenicity, allowing for effective binding to TNFα and treatment of TNFα-mediated disorders.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If conventional anti-TNFα antibodies are used, then TNFα binding activity is achieved, but stability and solubility are insufficient leading to aggregation
Solution Approach 1:
The patent applies parameter changes by modifying amino acid sequences at specific positions (e.g., position 42 in VL domain, position 89 in VH domain) to alter the physical-chemical properties of the antibody. These sequence modifications change the aggregation propensity and stability parameters of the antibody molecule, enabling improved solubility and reduced aggregation while maintaining TNFα binding activity.
Solution Approach 2:
The patent implements local quality by making targeted amino acid substitutions at specific local positions within the antibody variable domains rather than global modifications. The modifications focus on critical interfaces and regions prone to aggregation (such as the VL-VH interface and CDR regions), leaving other functional regions unchanged to preserve binding activity while improving local stability properties.
2Quantity of substance
If antibody concentration is increased for continuous treatment, then therapeutic efficacy is improved, but aggregation increases reducing solubility
Solution Approach 1:
The patent changes the aggregation parameter of the antibody through amino acid sequence modification, allowing the antibody to maintain stability at higher concentrations. The modified sequences reduce intermolecular aggregation interactions, enabling formulation at higher antibody concentrations necessary for continuous treatment without precipitating or aggregating.
Solution Approach 2:
The patent converts the harmful aggregation tendency into a beneficial property by selecting amino acid substitutions that specifically reduce aggregation propensity. The modifications at positions prone to aggregation (such as hydrophobic patches and interface regions) transform the antibody from an aggregation-prone molecule into a stable, soluble therapeutic suitable for high-concentration formulations.
3Stability of the object's composition
If antibody modifications are made to reduce aggregation, then stability is improved, but binding affinity to TNFα may be affected
Solution Approach 1:
The patent carefully selects modification sites that are local to aggregation-prone regions but distinct from the TNFα binding interface. By modifying specific positions in the framework regions and non-critical CDR positions while preserving the antigen-binding CDRs, the patent achieves local improvement in solubility without affecting the global binding function.
Solution Approach 2:
The patent applies partial action by making limited, targeted amino acid substitutions rather than extensive modifications. The modifications are applied selectively at key positions (such as one or two critical residues per variable domain) sufficient to reduce aggregation, avoiding excessive changes that could disrupt binding affinity or alter the antibody's functional properties.
Data Source
Figure 1
Figure 2A~2B
Figure 3A~3B
AI summary
The invention provides antibodies that are modified to reduce aggregration propensity, and methods of producing such antibodies. The present invention also provides particularly stable and soluble scFv antibodies and Fab fragments specific for TNF, which comprise specific light chain and heavy chain sequences that are optimized for stability, solubility, in vitro and in vivo binding of TNF, and low immunogenicity. The nucleic acids, vectors and host cells for expression of the recombinant antibodies of the invention, methods for isolating them and the use of said antibodies in medicine are also disclosed.