Amphipathic Polypeptide Enhances Soluble Antibody Fragment Expression
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Solution Overview
Problem
The challenge lies in converting full immunoglobulin molecules into antibody fragments that exhibit insufficient soluble recombinant expression, hindering the use of antibody fragments as therapeutic agents due to issues with expressibility and the need for alternative immunoglobulins, which can be costly and time-consuming to develop.
Innovation Solution
A method involving the combination of nucleic acid molecules encoding antibody variable regions with a diverse population, introduced into display systems like phage or prokaryotic systems, and the use of an N-terminal, cis-acting amphipathic polypeptide moiety to enhance soluble recombinant expression of antibody fragments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If direct conversion of parent immunoglobulin molecule into antibody fragment is performed, then antigen binding specificity is maintained, but soluble recombinant expression becomes insufficient
Solution Approach 1:
The patent introduces a soluble dimerization module as an intermediary component that mediates between the antibody variable regions and the expression system. This module acts as a bridge that enables soluble recombinant expression of antibody fragments that would otherwise be insoluble, while maintaining their antigen binding specificity through the Fv dimerization capability
Solution Approach 2:
The patent creates composite antibody fragment structures by combining antibody variable regions (VH and VL) with a soluble dimerization module. This composite construction integrates two functional elements: the antigen binding capability from the variable regions and the soluble expression capability from the dimerization module, thereby resolving the contradiction between maintaining specificity and achieving solubility
2Reliability
If antibody fragments with insufficient soluble expression are used, then therapeutic efficacy is reduced, but development of alternative immunoglobulins is required
Solution Approach 1:
The patent applies preliminary action by pre-designing and incorporating a soluble dimerization module into the antibody fragment construct before expression. This preliminary structural preparation ensures that the antibody fragments will be soluble and therapeutically effective without requiring time-consuming development of alternative immunoglobulins, thus resolving the contradiction between therapeutic efficacy and development time
3Ease of manufacture
If full immunoglobulin molecules are used instead of antibody fragments, then soluble expression is maintained, but tissue penetration and therapeutic efficacy are reduced
Solution Approach 1:
The patent applies segmentation by dividing the full immunoglobulin molecule into smaller antibody fragment components (VH and VL regions) while incorporating a soluble dimerization module. This segmentation reduces the overall size for better tissue penetration while the dimerization module maintains soluble expression capability, thereby resolving the contradiction between soluble expression and tissue penetration
Data Source
AI summary
The invention relates to methods of preparing antibody fragments. The invention further relates to antibody fragments prepared by these methods. The invention further relates to antibody variable regions comprised in antibody fragments producible by these methods.