DVD Binding Proteins for Homogeneous Bispecific Antibody Production
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Solution Overview
Problem
Current methods for producing bispecific antibodies face challenges such as molecular heterogeneity, reduced production yields, and increased immunogenicity due to the need for sophisticated purification procedures and site-directed modifications, which affect their stability and efficacy in binding multiple antigens.
Innovation Solution
Development of Dual Variable Domain (DVD) binding proteins with specific linker sequences that connect variable domains to form multivalent binding proteins capable of binding two or more antigens, including those like Prostaglandin E2 and Tumor Necrosis Factor alpha, using a combination of variable domains and constant domains to enhance antigen binding affinity and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If quadroma technology is used to produce bispecific antibodies, then two different antigen specificities can be achieved, but molecular heterogeneity increases and production yield decreases
Solution Approach 1:
The antibody is divided into separate variable domains (VH and VL) that can be independently arranged and paired. The patent uses segmented chain structures where VH1-VL1 and VH2-VL2 pairs are formed through controlled linkage, ensuring specific antigen binding while maintaining molecular homogeneity by preventing random pairing.
Solution Approach 2:
Linker peptides serve as intermediaries to connect variable domains in a controlled manner. The patent employs specific linker sequences (e.g., (GGGGS)n) that mediate the association between VH and VL domains, ensuring proper pairing and reducing molecular heterogeneity while maintaining the ability to bind multiple antigens.
2Adaptability or versatility
If chemical conjugation methods are used to produce bispecific antibodies, then antigen binding capability is achieved, but molecular homogeneity decreases and purification complexity increases
Solution Approach 1:
The patent replaces chemical conjugation methods with recombinant DNA-based expression systems. Instead of using chemical crosslinkers to attach different antibody fragments, the invention uses genetically engineered fusion proteins where variable domains are connected by peptide linkers, enabling production through standard recombinant expression and simplifying purification.
3Manufacturing precision
If site-directed modifications are made to produce homogeneous bispecific antibodies, then molecular homogeneity improves, but immunogenicity increases and stability decreases
Solution Approach 1:
The patent optimizes linker parameters (length, composition, flexibility) to achieve the right balance between homogeneity and stability. By adjusting linker properties such as using (GGGGS)n repeats with specific n values, the invention maintains molecular homogeneity while preserving antibody stability and reducing immunogenicity through natural amino acid sequences.
4Adaptability or versatility
If tandem scFv molecules are constructed to produce soluble bispecific antibodies, then antigen binding is achieved, but production yield decreases due to aggregation
Solution Approach 1:
The patent applies local quality control by introducing specific features at the linker regions between variable domains. The use of solubility-enhancing sequences (e.g., glycine-serine repeats) and optimized linker lengths at these critical locations prevents aggregation while maintaining soluble antigen binding capability, thereby improving production yield.
Data Source
AI summary
The present invention relates to engineered multivalent and multispecific binding proteins, methods of making, and specifically to their uses in the prevention, diagnosis, and/or treatment of disease.