Low Density Microarray for Influenza HA Protein Quantification
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Solution Overview
Problem
Current methods for quantifying influenza HA proteins and anti-influenza antibodies, such as SRID, ELISA, and HI assays, are time-consuming, costly, and complex, leading to inefficiencies and uncertainties in vaccine production and efficacy evaluation.
Innovation Solution
An array-based method using capture agents immobilized at various concentrations to generate binding curves, allowing for the quantification of unknown concentrations of target materials with reduced need for serial dilutions and enabling multiplex analysis for simultaneous quantification of multiple targets.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If single radial immunodiffusion (SRID) assay is used for quantification of influenza HA proteins, then measurement precision is improved, but loss of time and productivity deteriorate significantly
Solution Approach 1:
The patent segments the quantification process by immobilizing capture agents at multiple predetermined concentrations across different regions of the array substrate. This allows parallel measurement of multiple samples at different concentrations simultaneously, reducing the time required compared to sequential SRID assays while maintaining measurement precision through the calibrated concentration gradient.
Solution Approach 2:
The patent transitions from the traditional two-dimensional SRID diffusion method to a spatially encoded array format where concentration information is embedded in the spatial position of capture agents on the substrate. This dimensional reorganization enables simultaneous multi-point calibration curves to be generated in a single assay, dramatically reducing measurement time while preserving accuracy.
2Measurement precision
If SRID assay is performed with multiple serial dilutions to generate calibration curve, then measurement precision is improved, but device complexity and ease of operation worsen
Solution Approach 1:
The patent applies preliminary action by pre-immobilizing capture agents at multiple predetermined concentrations during array fabrication. This eliminates the need for researchers to perform time-consuming serial dilutions in the laboratory, as the concentration gradient is already established on the substrate. The calibration curve can be generated directly from the array signal intensities without additional manual dilution steps.
Solution Approach 2:
The array substrate serves itself by containing all necessary calibration information through the spatially distributed capture agents at known concentrations. When samples are applied, the system automatically generates the calibration curve data through the differential binding signals from the various capture agent concentrations, eliminating the need for external dilution operations and reducing procedural complexity.
3Measurement precision
If traditional ELISA or SRID methods are used, then quantification can be achieved, but cost and time consumption increase
Solution Approach 1:
The patent merges multiple functions into a single array substrate: calibration standards, sample analysis, and reference controls are all integrated on one platform. This consolidation eliminates the need for separate reagent preparations and multiple assay plates required by traditional ELISA or SRID methods, significantly reducing overall reagent consumption while maintaining quantification accuracy through the integrated design.
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
This approach significantly reduces costs and time by enabling rapid, accurate quantification of influenza HA proteins and antibodies, improving vaccine production efficiency and efficacy evaluation with improved sensitivity and ease of use compared to existing methods.
Implementation Method 1
capture agents being affixed to the array at a series of decreasing concentrations. In some embodiments, serial dilutions of a reference material are introduced, each dilution being contacted with a separate identical array. The reference material within each solution then binds to the corresponding capture agents on the array
Data Source
AI summary
Methods for the quantification of influenza HA proteins and anti-influenza antibodies for the fields of vaccine-related protein quantification, potency determination, and efficacy evaluation are provided. According to the technology, quantification is achieved by providing capture agents attached to an array in a series of decreasing concentrations. Serial dilutions of a reference material also may be introduced. The reference material within each solution binds to the capture agents on the array and is labeled with a label agent capable of producing a detectable signal used to construct a calibration curve. A target material of unknown concentration is introduced to a separate identical array, and the target material binds to the capture agents and also is labeled by a label agent to produce a detectable signal. The calibration curve based on the reference material is then utilized to determine the concentration of the target material without the need to perform replicate experiments.