Hypothesis: Protein nanoparticles have attracted increased interest due to their broad applications ranging from drug delivery and vaccines to biocatalysts and biosensors. The morphology and the size of the nanoparticles play a crucial role in determining their suitability for different applications. Yet, effectively controlling the size of the nanoparticles is still a significant challenge in their manufacture. The hypothesis of this paper is that the assembly conditions and size of protein particles can be tuned via a mechanical route by simply modifying the mixing time and strength, while keeping the chemical parameters constant. Experimental: We use an acoustically actuated, high throughput, ultrafast, microfluidic mixer for the assembly of protein particles with tuneable sizes. The performance of the acoustic micro-mixer is characterized via Laser Doppler Vibrometry and image processing. The assembly of protein nanoparticles is monitored by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Findings: By changing actuation parameters, the turbulence and mixing in the microchannel can be precisely varied to control the initiation of protein particle assembly while the solution conditions of assembly (pH and ionic strength) are kept constant. Importantly, mixing times as low as 6 ms can be achieved for triggering protein assembly in the microfluidic channel. In comparison to the conventional batch process of assembly, the acoustic microfluidic mixer approach produces smaller particles with a more uniform size distribution, promising a new way to manufacture protein particles with controllable quality. (C) 2020 Elsevier Inc. All rights reserved.