BTN2A1-Binding γδ TCR Peptides for Higher Affinity and Stability
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Solution Overview
Problem
Existing strategies for enhancing the affinity and stability of Vγ9Vδ2 TCRs for BTN2A1 interaction are inadequate, leading to suboptimal potency in tumor recognition and clearance, and production yield is limited, hindering the effectiveness of γδT cell-based therapeutics.
Innovation Solution
Identification of mutant BTN2A1 binding peptides in the Vγ9 domain, such as Vγ9E22W, Vγ9T91H, and Vγ9R73Y, which enhance affinity and stability, respectively, allowing for more potent γδT cell receptor-based therapeutic agents and improved production yields.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type Vγ9Vδ2 TCRs are used for tumor recognition, then the system is simple and production is easy, but the affinity for BTN2A1 is insufficient (40-50 μM) leading to suboptimal tumor clearance potency
Solution Approach 1:
The patent applies parameter changes by mutating specific amino acid residues in the Vγ9 domain (positions 22, 73, and 91) to alter the binding parameters of the TCR-BTN2A1 interaction. The E22W mutation increases affinity by 10-fold, while R73Y and T91H mutations enhance stability, transforming the TCR from low-affinity wild-type to high-affinity mutant with improved tumor clearance potency.
2Reliability
If affinity enhancement mutations are introduced to improve tumor recognition, then binding affinity increases, but production yield is limited and manufacturing becomes more difficult
Solution Approach 1:
The patent simultaneously optimizes multiple parameters by introducing the R73Y mutation which enhances protein stability and the T91H mutation that further improves binding characteristics. These parameter changes in the amino acid sequence enable both high binding affinity (10-fold increase) and improved production yield, resolving the contradiction between affinity enhancement and manufacturability.
3Reliability
If multiple mutations are introduced to enhance both affinity and stability, then therapeutic effectiveness improves, but the complexity of characterization and quality control increases
Solution Approach 1:
The patent applies local quality by introducing targeted point mutations at specific positions (22, 73, 91) rather than global modifications. Each mutation has a localized function: E22W for affinity enhancement, R73Y for stability improvement, and T91H for additional binding optimization. This localized approach improves therapeutic effectiveness while maintaining manageable characterization complexity through focused analysis of specific residue changes.
Data Source
AI summary
Disclosed is a Butyrophilin Subfamily 2 Member A1 (BTN2A1) binding peptide with an amino acid sequence comprising an amino acid other than glutamic acid, for example, tryptophan, at a position corresponding to position 22 shown in SEQ ID NO: 1; an amino acid other than arginine, for example, tyrosine, at a position corresponding to position 73 shown in SEQ ID NO: 1; and/or an amino acid other than threonine, for example, histidine or tryptophan, at a position corresponding to position 81 shown in SEQ ID NO: 1. The BTN2A1 binding peptide may be comprised in an (exogeneous) immune receptor or extracellular domain thereof, preferably a γδ T-cell receptor or extracellular domain thereof, more preferably a γ9δ2 T-cell receptor or extracellular domain thereof. This disclosure particularly relates to use in therapy, preferably for use in treatment of a cancer and/or an infection, or for use in diagnostic methods.