Design and implement a combination of laser force cytology cell analysis and sorting and capillary electrophoresis (CE)-based methods to screen for viral and bacterial contamination, along other process parameters.
Laser force cytology gives virus detection sensitivity equal to or exceeding that of cytopathic effect (CPE) assays.
LFC instrument installed for detection of MMV in CHO cells – also found sensitivity equal to or exceeding CPE using statistical methods developed in the project.
Micelle-tagging electrophoresis (MTE), using peptide nucleic acid (PNA) probes, detected viral DNA / RNA down to 10 fM in a 10-minute run.
Rapid at-line detection of viral contaminants
Allows for process troubleshooting and screening of raw materials, cell banks and bulk harvest materials
Publication: P.R. Hayes. T.M. Przybycien, J.W. Schneider. ?Viral adventitious agent detection using laser force cytology: Intrinsic cell property changes with infection and comparison to in vitro testing,? submitted to Biotech. Bioeng. (2021) Direct, PCR-less Tagging of Viral RNA using Micelle Tagging Electrophoresis (MTE) - June 19, 2019
Hayes, P., Przybycien, T., & Schneider, J., Viral adventitious agent detection using laser force cytology: Intrinsic cell property changes with infection and comparison to in vitro testing, Biotechnology and Bioengineering, DOI:10.1002/bit.27957
Hayes, P., Detection of Viruses Using Biophysical Properties of Infected Cells for Biologics and Vaccines Testing, Carnegie Mellon University, Pittsburgh, PA, August 1, 2022. https://doi.org/10.1184/R1/22057130
Hui, K., High Resolution Nucleic Acid Electrophoresis by Snagging Mechanisms in Wormlike Micelle Networks, Carnegie Mellon University, Pittsburgh, PA, August 1, 2022. https://www.proquest.com/openview
Schneider, J.W., Przybycien, Presenter, Gordon Research Conference: Biotherapeutics and Vaccines Development, Galveston, TX, January 6, 2019.
Schneider, J.W., Przybycien, T.M., Hart, S., Hebert, C., Elvington, S., & St. John, R., Presenter, QSP Project: At-line Detection of Viral and Bacterial Contaminants in Mammalian Cell Culture using High Affinity Probes and Label-free Single-cell Analysis, Summit, NJ, May 17, 2018.
At-line Detection of Viral and Bacterial Contaminants in Mammalian Cell Culture Using High Affinity Probes and Label-free Single-cell Analysis (QSP1.0-036), NIIMBL Member Forum, Virtual, January 24, 2019.
Schneider, J. W., Hui, K., Yan, L., & Przybycien, T. M., Direct, PCR-less Tagging of Viral RNA using Micelle Tagging Electrophoresis (MTE), 93rd ACS Colloid and Surface Science Symposium, Atlanta, GA, June 1, 2019.
Schneider, J.W., Przybycien, T.M., Adventitious agent detection in continuous cell culture – What can we learn from sensing in biomedical and biowarfare applications?, Boston, MA, April 6, 2018.
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Carnegie Mellon University
Genentech, Inc.
LumaCyte
