Rapid ELISA Assay Using Quick Coating Buffer
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Solution Overview
Problem
Conventional ELISA methods are time-consuming, requiring several hours for antigen detection and quantification, necessitating a need for a rapid and efficient process.
Innovation Solution
The development of a rapid enzyme-linked immunosorbent assay (ELISA) method using a quick coating buffer and blocking reagents, reducing coating and blocking times to as little as five minutes, and integrating multiple incubation steps into fewer, more efficient processes, such as simultaneous incubation with multiple detection agents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional ELISA methods are used with standard coating and blocking procedures, then reliable antigen detection is achieved, but the assay time is excessively long (several hours)
Solution Approach 1:
The patent changes the chemical parameters of the coating buffer by adding organic solvents (DMSO, ethanol, isopropanol) to conventional buffers. This parameter modification enables rapid antigen coating within 5 minutes while maintaining detection accuracy, resolving the contradiction between reliable detection and excessive assay time
Solution Approach 2:
The patent performs preliminary optimization of coating and blocking conditions using the quick-coating buffer before the actual assay. This preliminary action establishes optimal conditions that enable rapid subsequent assays, reducing overall assay time without compromising detection reliability
2Reliability
If multiple incubation steps are performed sequentially as in conventional ELISA, then thorough detection is achieved, but the process becomes time-consuming and complex
Solution Approach 1:
The patent merges multiple sequential incubation steps into a single simultaneous incubation step where primary and secondary antibodies are incubated together. This merging maintains detection reliability through proper reagent ratios while significantly improving productivity by reducing the number of separate steps
Solution Approach 2:
The patent develops a universal quick-coating buffer formulation that works across different ELISA formats (direct, indirect, sandwich). This multi-functional buffer system maintains reliable detection across various assay types while enabling rapid processing, thus improving productivity without sacrificing reliability
3Manufacturing precision
If standard coating buffers are used with extended incubation times, then complete antigen coating is achieved, but the coating step becomes unnecessarily long
Solution Approach 1:
The patent modifies the chemical composition parameters of the coating buffer by incorporating organic solvents at optimized concentrations. These parameter changes increase the coating kinetics, achieving complete antigen coating in 5 minutes compared to the conventional 2+ hours, thus resolving the contradiction between coating completeness and coating time
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 the overall time required for ELISA assays from several hours to under two hours, enhancing detection efficiency and cost-effectiveness while maintaining accurate results.
Implementation Method 1
coating a solid phase with an antigen dissolved in a quick coating buffer
Implementation Method 2
blocking the solid phase with a blocking reagent dissolved in a quick blocking buffer for between about two to ten minutes
Data Source
AI summary
The present invention provides improved and rapid detection methods for an antigen such as a chemical compound, a peptide, a nucleic acid, or a protein released from cells or virus particles in situ. The detection time for an antigen can be dramatically reduced relative to conventional technologies. The technology can particularly be used, for example, to modify and reduce the detection time significantly in traditional ELISA, and also Western blot or Dot blot assays. The improved ELISA method is rapid, economical, reproducible, simple and automatable. Also provided are compositions and kits for using the improved ELISA methods for the rapid detection of antigens.