Measurements were carried out on a Bruker UltrafleXtreme MALDI-TOF mass spectrometer (Bruker Daltonics, Bremen, Germany). First, the buffer and salt components from the samples of PVY virus and VLP constructs were exchanged with pure water, and samples were concentrated to 1.5 to 3.0 mg/ml with centrifugation through 10-kDa cutoff membranes. Samples were then mixed with saturated solution of sinapinic acid in a mixture of acetonitrile and 0.1% trifluoroacetic acid (1:1, v/v) in a 1:1 (v/v). One microliter of this solution was spotted on the target plate (dried-droplet method). The linear positive ion mode was used to acquire the mass spectra of the samples. The calibration was done externally using protein calibration standards I and II (Bruker Daltonics). Sample preparation procedure for the standards was the same as that for other samples. The standards were spotted on the nearest neighboring positions.

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