TY - JOUR
T1 - On-Chip Acousto Thermal Shift Assay for Rapid and Sensitive Assessment of Protein Thermodynamic Stability
AU - Ding, Yonghui
AU - Ball, Kerri A.
AU - Webb, Kristofor J.
AU - Gao, Yu
AU - D'Alessandro, Angelo
AU - Old, William M.
AU - Stowell, Michael H.B.
AU - Ding, Xiaoyun
N1 - Funding Information:
The ATSA devices were fabricated in JILA clean room at University of Colorado Boulder. This work was supported by startup funding of University of Colorado Boulder to X.D. and CU Boulder MCDB Neurodegenerative Disease Fund to M.H.B.S., and DARPA cooperative agreement, 13‐34‐RTA‐FP‐007, to W.M.O. and M.H.B.S.
Publisher Copyright:
© 2020 Wiley-VCH GmbH
PY - 2020/10/1
Y1 - 2020/10/1
N2 - 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 interactions. However, existing TSAs have limitations, such as being time consuming, labor intensive, or having low sensitivity. Herein, an acousto thermal shift assay (ATSA), the first ultrasound enabled TSA, is reported for real-time analysis of protein thermodynamic stability. It capitalizes the coupling of unique acoustic mechanisms to achieve protein unfolding, concentration, and measurement on a single microfluidic chip within minutes. Compared to conventional TSA methods, the ATSA technique enables ultrafast (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.
AB - 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 interactions. However, existing TSAs have limitations, such as being time consuming, labor intensive, or having low sensitivity. Herein, an acousto thermal shift assay (ATSA), the first ultrasound enabled TSA, is reported for real-time analysis of protein thermodynamic stability. It capitalizes the coupling of unique acoustic mechanisms to achieve protein unfolding, concentration, and measurement on a single microfluidic chip within minutes. Compared to conventional TSA methods, the ATSA technique enables ultrafast (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.
KW - lab on a chip
KW - microfluidics
KW - pharmaceutical diagnostics, molecular diagnostics
KW - protein thermal shift assay
KW - surface acoustic waves
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U2 - 10.1002/smll.202003506
DO - 10.1002/smll.202003506
M3 - Article
C2 - 32893496
AN - SCOPUS:85090306641
SN - 1613-6810
VL - 16
JO - Small
JF - Small
IS - 41
M1 - 2003506
ER -