Microfluidic deposition for resolving single-molecule protein architecture and heterogeneity

Francesco Simone Ruggeri, Jerome Charmet, Tadas Kartanas, Quentin Peter, Sean Chia, Johnny Habchi, Christopher M. Dobson, Michele Vendruscolo and Tuomas P. J. Knowles ()
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Francesco Simone Ruggeri: University of Cambridge
Jerome Charmet: University of Cambridge
Tadas Kartanas: University of Cambridge
Quentin Peter: University of Cambridge
Sean Chia: University of Cambridge
Johnny Habchi: University of Cambridge
Christopher M. Dobson: University of Cambridge
Michele Vendruscolo: University of Cambridge
Tuomas P. J. Knowles: University of Cambridge

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: Abstract Scanning probe microscopy provides a unique window into the morphology, mechanics, and structure of proteins and their complexes on the nanoscale. Such measurements require, however, deposition of samples onto substrates. This process can affect conformations and assembly states of the molecular species under investigation and can bias the molecular populations observed in heterogeneous samples through differential adsorption. Here, we show that these limitations can be overcome with a single-step microfluidic spray deposition platform. This method transfers biological solutions to substrates as microdroplets with subpicoliter volume, drying in milliseconds, a timescale that is shorter than typical diffusion times of proteins on liquid–solid interfaces, thus avoiding surface mass transport and change to the assembly state. Finally, the single-step deposition ensures the attachment of the full molecular content of the sample to the substrate, allowing quantitative measurements of different molecular populations within heterogeneous systems, including protein aggregates.

Date: 2018
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DOI: 10.1038/s41467-018-06345-4

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