Optical tweezers-controlled hotspot for sensitive and reproducible surface-enhanced Raman spectroscopy characterization of native protein structures

Xin Dai, Wenhao Fu, Huanyu Chi, Vince St. Dollente Mesias, Hongni Zhu, Cheuk Wai Leung, Wei Liu () and Jinqing Huang ()
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Xin Dai: The Hong Kong University of Science and Technology
Wenhao Fu: The Hong Kong University of Science and Technology
Huanyu Chi: The Hong Kong University of Science and Technology
Vince St. Dollente Mesias: The Hong Kong University of Science and Technology
Hongni Zhu: The Hong Kong University of Science and Technology
Cheuk Wai Leung: The Hong Kong University of Science and Technology
Wei Liu: The University of Hong Kong
Jinqing Huang: The Hong Kong University of Science and Technology

Nature Communications, 2021, vol. 12, issue 1, 1-9

Abstract: Abstract Surface-enhanced Raman spectroscopy (SERS) has emerged as a powerful tool to detect biomolecules in aqueous environments. However, it is challenging to identify protein structures at low concentrations, especially for the proteins existing in an equilibrium mixture of various conformations. Here, we develop an in situ optical tweezers-coupled Raman spectroscopy to visualize and control the hotspot between two Ag nanoparticle-coated silica beads, generating tunable and reproducible SERS enhancements with single-molecule level sensitivity. This dynamic SERS detection window is placed in a microfluidic flow chamber to detect the passing-by proteins, which precisely characterizes the structures of three globular proteins without perturbation to their native states. Moreover, it directly identifies the structural features of the transient species of alpha-synuclein among its predominant monomers at physiological concentration of 1 μM by reducing the ensemble averaging. Hence, this SERS platform holds the promise to resolve the structural details of dynamic, heterogeneous, and complex biological systems.

Date: 2021
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DOI: 10.1038/s41467-021-21543-3

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