On-chip Acousto Thermal Shift Assay for Rapid and Sensitive Assessment of Protein Thermodynamic Stability

TL;DR

This paper introduces a novel ultrasound-enabled on-chip thermal shift assay (ATSA) that significantly accelerates and enhances the sensitivity of protein stability and interaction measurements, enabling rapid, label-free analysis.

Contribution

The paper presents the first ultrasound-based TSA, coupling acoustic mechanisms with microfluidics for real-time, ultra-fast, and highly sensitive protein stability assessment.

Findings

ATSA is at least 30 times faster than conventional methods.

It offers 7-34 fold higher sensitivity.

The technique enables label-free, real-time analysis of protein interactions.

Abstract

Thermal shift assays (TSAs) have been extensively used to study thermodynamics of proteins and provide an efficient means to assess protein-ligand binding or protein-protein interaction. However, existing TSAs have limitations such as time consuming, labor intensive, or low sensitivity. Here we introduce a novel acousto thermal shift assay (ATSA), the first ultrasound enabled TSA, for real-time analysis of protein thermodynamic stability. It capitalizes the novel coupling of unique acoustic mechanisms to achieve protein unfolding, concentration, and measurement on a single microfluidic chip within minutes. Compared to conventional TSA methods, our ATSA technique enabled ultra-fast (at least 30 times faster), highly sensitive (7-34 folds higher), and label-free monitoring of protein-ligand interactions and protein stability. ATSA paves new avenues for protein analysis in biology, medicine and fast diagnosis.