PFAS Protein Binding Affinity Screening Using Thermal Shift Fluorescence

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Solution Overview

Problem

There is a lack of high-throughput and predictive methods to characterize protein binding affinities for a wide range of per- and polyfluoroalkyl substances (PFAS), due to limited data on their physiochemical and toxicokinetic properties, which hinders the evaluation of their bioactivity and potential toxicity.

Innovation Solution

A method involving differential scanning fluorimetry to determine melting temperatures (Tm) of proteins in the presence and absence of PFAS, calculating ΔTm, and fitting the data to equations to determine dissociation constants, enabling rapid characterization of protein binding affinities.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If traditional methods are used to characterize protein binding for PFAS, then measurement precision may be adequate, but productivity is extremely low due to lack of high-throughput capabilities

Engineering Contradiction:

Improvethroughput of protein binding characterizationVSAvoidaccuracy of protein binding affinity data

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The patent replaces traditional mechanical/physical binding assays with a fluorescence-based optical detection system. The method uses environment-sensitive fluorophores that change fluorescence properties upon binding to proteins, enabling rapid, high-throughput measurement of protein binding affinities for multiple PFAS compounds simultaneously while maintaining measurement precision through optical detection rather than mechanical means.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the measurement parameter from traditional binding affinity measurements to melting temperature (Tm) shifts detected by fluorescence. By monitoring changes in protein thermal stability (ΔTm) caused by PFAS binding, the method enables high-throughput screening while preserving the ability to calculate accurate binding affinities through the relationship between Tm shifts and dissociation constants.

Inventive Principle:

Principle #35Parameter changes

2Loss of information

If comprehensive data collection for all PFAS is attempted using traditional methods, then complete characterization could be achieved, but loss of time would be excessive

Engineering Contradiction:

Improvecompleteness of PFAS physiochemical property dataVSAvoidtime required to characterize thousands of PFAS

Core Design Contradiction:

Loss of informationVSLoss of time

Solution Approach 1:

The patent creates a high-throughput screening platform that can rapidly generate binding affinity data for large numbers of PFAS compounds. By using standardized protein targets (such as serum albumin) and automated fluorescence detection, the method produces comprehensive datasets for thousands of PFAS in a fraction of the time required by traditional methods, effectively copying the essential binding information across many compounds simultaneously.

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent develops a universal assay platform that can screen multiple classes of PFAS compounds against various protein targets using the same basic methodology. The environment-sensitive fluorophore approach and Tm shift measurement can be applied to diverse PFAS structures (carboxylic acids, sulfonic acids, ethers, etc.) and different proteins, enabling comprehensive data collection across the entire PFAS chemical space through a single multi-functional system.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Productivity

If predictive computational strategies are used for PFAS characterization, then productivity increases, but measurement precision is limited by lack of fundamental data

Engineering Contradiction:

Improvespeed of PFAS property predictionVSAvoidaccuracy of predictive models

Core Design Contradiction:

ProductivityVSMeasurement precision

Solution Approach 1:

The patent performs preliminary experimental characterization of protein binding affinities for numerous PFAS compounds using the high-throughput fluorescence method. These experimentally determined binding constants serve as preliminary data that can be used to train and validate predictive computational models, thereby improving the accuracy of future predictions while maintaining high productivity through the established screening platform.

Inventive Principle:

Principle #10Preliminary action

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

Enables rapid, high-throughput determination of protein binding properties for PFAS, overcoming limitations of existing methods by providing scalable and accurate data on protein binding affinities.

Implementation Method 1

providing a first composition comprising a polyfluoroalkyl substance, an environment-sensitive fluorophore, a protein, and an aqueous buffer solution, and determining a melting temperature (Tm) for the protein in the first composition

Methodology Applied

Scientific EffectFluorescence: Fluorescence