Nanobody Engineering for TNF-alpha Affinity and Stability
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Solution Overview
Problem
Current Nanobodies against TNF-alpha have limitations in terms of affinity, stability, specificity, and immunogenicity, which affect their therapeutic and diagnostic efficacy for diseases mediated by TNF-alpha.
Innovation Solution
Development of improved Nanobodies with enhanced affinity, stability, and specificity, including multispecific formats that bind to specific epitopes on TNF-alpha, such as those comprising amino acid residues Gln at position 88, Lys at position 90, and Glu at position 146, and linked in a manner allowing intramolecular binding to TNF receptor sites, reducing receptor crosslinking and signal transduction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional Nanobodies against TNF-alpha are used, then they can bind to TNF-alpha, but their affinity, stability, and specificity are insufficient for optimal therapeutic efficacy
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the Nanobody, specifically optimizing framework regions and complementarity determining regions to enhance binding affinity, thermal stability, and specificity for TNF-alpha while maintaining the ability to block receptor crosslinking
Solution Approach 2:
The patent creates composite structures by combining multiple functional elements within the Nanobody sequence, including stabilized framework regions and optimized CDRs, to achieve enhanced overall performance in terms of affinity, stability, and therapeutic efficacy
2Power
If Nanobodies are designed to bind with high affinity to TNF-alpha, then potency increases, but immunogenicity may increase reducing therapeutic suitability
Solution Approach 1:
The patent applies parameter changes by humanizing the Nanobody sequence and optimizing specific amino acid positions to reduce immunogenicity while preserving or enhancing binding potency against TNF-alpha, making the molecule more suitable for therapeutic applications in human patients
3Reliability
If Nanobodies bind to TNF-alpha epitopes, then they can inhibit receptor crosslinking, but their stability and half-life in vivo may be insufficient
Solution Approach 1:
The patent applies parameter changes by engineering the Nanobody sequence to enhance structural stability and resistance to proteolysis, thereby extending the half-life in vivo while maintaining the ability to effectively inhibit TNF receptor crosslinking and signal transduction
Data Source
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AI summary
The present invention relates to improved Nanobodies™ against Tumor Necrosis Factor-alpha (TNF-alpha), as well as to polypeptides comprising or essentially consisting of one or more of such Nanobodies. The invention also relates to nucleic acids encoding such Nanobodies and polypeptides; to methods for preparing such Nanobodies and polypeptides; to host cells expressing or capable of expressing such Nanobodies or polypeptides; to compositions comprising such Nanobodies, polypeptides, nucleic acids or host cells; and to uses of such Nanobodies, such polypeptides, such nucleic acids, such host cells or such compositions, in particular for prophylactic, therapeutic or diagnostic purposes, such as the prophylactic, therapeutic or diagnostic purposes.