Microfluidic affinity selection of active SARS-CoV-2 virus particles
Sachindra S. Gamage, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Thilanga N. Pahattuge, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Harshani Wijerathne, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Katie Childers, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Swarnagowri Vaidyanathan, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Uditha S. Athapattu, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Lulu Zhang, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Zheng Zhao, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Mateusz L. Hupert, BioFluidica Inc., San Diego, CA 92121, USA.
Rolf M. Muller, BioFluidica Inc., San Diego, CA 92121, USA.
Judy Muller-Cohn, BioFluidica Inc., San Diego, CA 92121, USA.
Janet Dickerson, BioFluidica Inc., San Diego, CA 92121, USA.
Dylan Dufek, BioFluidica Inc., San Diego, CA 92121, USA.
Brian V. Geisbrecht, Department of Biochemistry and Molecular Biophysics, Kansas State University, Manhattan, KS 66506, USA.
Harsh Pathak, Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Ziyan Pessetto, Sinochips Diagnostics, Olathe, KS 66061, USA.
Gregory N. Gan, Department of Radiation Oncology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Junseo Choi, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Sunggook Park, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Andrew K. Godwin, Center of BioModular Multiscale Systems for Precision Medicine, The University of Kansas, Lawrence, KS 66045, USA.
Malgorzata A. Witek, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Steven A. Soper, Department of Chemistry, The University of Kansas, Lawrence, KS 66045, USA.
Abstract
We report a microfluidic assay to select active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles (VPs), which were defined as intact particles with an accessible angiotensin-converting enzyme 2 receptor binding domain (RBD) on the spike (S) protein, from clinical samples. Affinity selection of SARS-CoV-2 particles was carried out using injection molded microfluidic chips, which allow for high-scale production to accommodate large-scale screening. The microfluidic contained a surface-bound aptamer directed against the virus's S protein RBD to affinity select SARS-CoV-2 VPs. Following selection (~94% recovery), the VPs were released from the chip's surface using a blue light light-emitting diode (89% efficiency). Selected SARS-CoV-2 VP enumeration was carried out using reverse transcription quantitative polymerase chain reaction. The VP selection assay successfully identified healthy donors (clinical specificity = 100%) and 19 of 20 patients with coronavirus disease 2019 (COVID-19) (95% sensitivity). In 15 patients with COVID-19, the presence of active SARS-CoV-2 VPs was found. The chip can be reprogrammed for any VP or exosomes by simply changing the affinity agent.
