Purification of Lymphocytes by Acoustic Separation in Plastic Microchannels

Abstract

Emerging cell therapies have created new demands for instruments that will increase processing efficiency. Purification of lymphocytes prior to downstream steps of gene transfer currently relies on centrifugal separation, which has drawbacks in output sample purity and process automation. Here, we present an alternative approach to blood cell purification using acoustic forces in plastic microchannels. We provide details regarding the system's ability to purify lymphocytes relative to other blood cell types while maintaining a high overall recovery, testing performance starting from leukapheresis product, buffy coat, and whole blood. Depending on settings, the device achieves for lymphocytes up to 97% purity and up to 68% recovery, and depletes 98% of monocytes while also reducing red cells and platelets. We expect that future scale-up of our system for increased throughput will enable its incorporation in the cell therapy workflow, and that it could ultimately reduce costs and expand access for patients.

Label-free separation of leukocyte subpopulations using high throughput multiplex acoustophoresis

“…Furthermore, acoustophoresis has been shown to be a gentle method which does not impact cell viability or functional capacity of the separated sample. 21,23,[27][28][29][30][31][32][33] There is a large size overlap between the different white blood cell populations with a median diameter (range) of 7.2 μm (5.5-10 μm) for lymphocytes, 9.5 μm (7.5-12 μm) for monocytes and 9.5 μm (8.5-11 μm) for granulocytes as determined by coulter counter measurements (Fig. S2 †).…”

Whole-System Ultrasound Resonances as the Basis for Acoustophoresis in All-Polymer Microfluidic Devices

“…However, partly due to the lack of good theoretical understanding, it has proven difficult to make good all-polymer BAW-devices for acoustofluidics. But a few results for such devices have been published, including focusing of polymer beads [32][33][34][35], lipids [32], and red blood cells [33,36], as well as blood-bacteria separation [37] and purification of lymphocytes [38].…”

Acoustic Microfluidic Separation Techniques and Bioapplications: A Review

“…However, to realize their full potential, there are some limitations still required to be overcome in future. For example, the throughput of acoustic microfluidic separation devices is relatively low in a single-channel microfluidic device [ 122 ]. Moreover, acoustic separation microfluidic devices usually need to integrate with bulky instruments, such as power supply, voltage amplifiers, and syringe pumps, which increases the complexity of the system.…”