Universal Capture Array for Multiplexed Influenza Protein Quantification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for protein quantification in influenza vaccines, such as SRID, are low-throughput, require frequent reagent updates, and cannot accurately measure degradation, making them inefficient and costly, especially for multivalent vaccines that need to account for seasonal mutations.
Innovation Solution
A universal capture array method that uses a microarray with spatially distinct spots of murine antibody capture agents targeting conformational and linear epitopes of hemagglutinin, allowing for multiplexed quantification of subcomponents without requiring reagent updates and enabling the measurement of protein degradation without a reference sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If SRID assay is used for protein quantification, then measurement precision is maintained, but productivity is severely limited and hands-on time is excessive
Solution Approach 1:
The assay is segmented into distinct functional zones on the microarray substrate: capture agent spots for antigen capture, detection agent spots for signal generation, and control spots for quality assurance. This spatial segmentation enables simultaneous processing of multiple samples and antigens, dramatically increasing throughput while maintaining the precision of individual measurements through dedicated functional areas.
Solution Approach 2:
The microarray platform serves multiple functions: it quantifies different influenza antigens simultaneously, identifies degradation products, and provides controls for assay validation. The universal capture agents can bind multiple antigen types, and the same microarray structure handles both quantification and degradation detection, eliminating the need for separate assays and reducing hands-on time.
2Measurement precision
If SRID assay is used for protein quantification, then measurement precision is maintained, but loss of time is excessive due to 2-3 day duration
Solution Approach 1:
Capture agents are pre-immobilized on the microarray substrate in specific spatial patterns before sample addition. This preliminary preparation eliminates the need for time-consuming assembly steps during the assay and allows samples to be processed immediately upon arrival, reducing the overall assay duration from 2-3 days to a significantly shorter timeframe while preserving measurement precision through pre-optimized capture conditions.
Solution Approach 2:
The microarray assay enables continuous processing where multiple detection reagents can be added sequentially without removing the substrate, and readings can be taken continuously as signals develop. This continuous action eliminates idle time between steps and allows the assay to reach completion much faster than the intermittent, multi-day SRID protocol while maintaining accurate quantification.
3Measurement precision
If SRID assay is used for protein quantification, then measurement precision is maintained, but device complexity increases due to multiple reagents and subjective readout
Solution Approach 1:
Detection agents are conjugated to fluorescent or colorimetric labels that produce distinct optical signals upon binding to captured antigens. This color/fluorescence change provides an objective, instrument-readable output that eliminates subjective visual interpretation while maintaining quantification accuracy. The spatially resolved signals from different spots can be automatically analyzed by imaging systems, reducing complexity in the readout process.
Solution Approach 2:
The microarray substrate acts as an intermediary platform that organizes and presents multiple capture and detection agents in a structured format. This intermediary structure simplifies the overall assay by providing a pre-arranged framework for reagent interactions, eliminating the need for complex manual assembly and interpretation procedures associated with traditional SRID methods.
4Adaptability or versatility
If universal capture agents are used, then adaptability to seasonal mutations is improved, but manufacturing precision is required to ensure consistent binding across diverse antigens
Solution Approach 1:
Different capture agents with specific binding characteristics are placed at different spatial locations on the microarray. Each capture agent is optimized for its local function, with properties tailored to bind specific antigen types or degradation products. This local optimization ensures consistent and reliable binding across diverse antigens while maintaining adaptability to seasonal variations through the diversity of capture agents available on the same platform.
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 method provides a high-throughput, cost-effective, and reliable means to quantify and subtype influenza proteins, reducing hands-on time and variability, and can accurately measure protein stability across different vaccine formulations and strains.
Implementation Method 1
The detection antibody is biotinylated and binds to the captured antigen
Implementation Method 2
streptavidin conjugated to a fluorescent reporter or other detectable label
Data Source
AI summary
A universal array for multiplexed quantification of variable hemagglutinin such as subcomponents of multivalent annual influenza vaccines that is robust to variations in proteins such as mutations and is capable of quantifying degradation of proteins. Universal capture array (100) comprises one or more substrates (102) and a low-density microarray (104) of sub-arrays (108) comprising spots (106a-c). The microarray (104) is contacted with one or more targets (202) at one or more unknown concentrations, and bound complexes (203) are formed and subsequently quantified with a suitable method. Quantified signals are compared to calibration curves to obtain one or more unknown concentrations and/or quantify degradation of the one or more targets (202). Other embodiments are described and shown.