pH-Gradient SPR Assay for Antibody Dissociation Kinetics
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Solution Overview
Problem
Existing SPR technologies struggle to effectively determine pH-dependent interactions of antibodies, limiting the optimization of antibody pharmacokinetics and neutralization efficiency, as they cannot accurately measure pH-dependent binding and dissociation kinetics.
Innovation Solution
A novel SPR-based assay that applies a pH-gradient to the measurement system, monitoring the pH-dependent binding assay, immobilizing the antibody on a solid phase, and the pH-dependent binding assay, immobilizing the antibody on a solid phase, and the pH-dependent binding assay, and the pH-dependent binding assay, immobilizing the antibody on a solid phase, and applying a pH-gradient to determine pH-dependent binding and dissociation kinetics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional SPR technology is used to measure antibody-antigen interactions, then binding constants can be determined, but pH-dependent interactions cannot be accurately measured
Solution Approach 1:
The patent applies a pH-gradient across the SPR measurement system, transitioning from uniform pH conditions to spatially varying pH conditions. This allows simultaneous measurement of antibody-antigen binding at different pH values, enabling accurate determination of pH-dependent interaction constants and dissociation kinetics that conventional single-pH SPR cannot measure.
2Reliability
If antibody affinity is increased to improve neutralization efficiency, then smaller antibody amounts are needed, but the complexity of affinity maturation increases
Solution Approach 1:
The patent replaces complex iterative affinity maturation processes with a simpler pH-dependent binding assessment method. By measuring binding characteristics across a pH-gradient, researchers can identify antibodies with optimal pH-dependent binding profiles that naturally provide enhanced neutralization efficiency without requiring multiple rounds of complex genetic engineering and selection.
3Duration of action of moving object
If antibody dose is increased to sustain neutralization effect, then neutralization duration is prolonged, but the cost and complexity of treatment increases
Solution Approach 1:
The patent enables identification of antibodies with optimized pH-dependent binding characteristics that maintain stable binding at physiological pH (prolonging duration of action) while allowing dissociation at acidic pH (enabling recycling). This allows achieving sustained neutralization effects with lower antibody doses by selecting antibodies whose binding properties are optimized across the pH-gradient, rather than increasing dose to extend duration.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The assay provides detailed insights into pH-dependent antibody-antigen interactions, enabling the selection and optimization of antibodies with improved pharmacokinetics and neutralization efficiency by identifying pH-dependent binding and dissociation kinetics.
Implementation Method 1
SPR (surface plasmon resonance) is a biosensor-based technology to measure real time protein-protein interaction
Data Source
AI summary
Herein is reported a method for determining the dissociation rate constant kct of an antibody from its antigen at the dissociation pH-value, wherein the method comprises the steps of immobilizing at a first pH-value the antibody on a solid phase to which the antigen of the antibody has been conjugated; applying a pH-gradient from the first pH-value to the dissociation pH-value and thereafter maintaining the pH-value at said dissociation pH-value; and recording the binding signal during the maintaining of the pH-value and calculating therefrom the dissociation rate constant ka of the antibody from its antigen at the dissociation pH-value.