title: Microscale acoustic disruption of mammalian cells for intracellular product release.
abstract: High-throughput microscale models for cell culture are critical for biopharmaceutical process development and drug discovery compound screening. While analytical methods are readily available for quantifying cell number and secreted product concentration, the recovery and measurement of intracellular products are significantly affected by the method of cell disruption. For example, the detergents often used in product extraction are incompatible with lipid-enveloped viruses. To provide an alternative to detergent-mediated disruption of mammalian cells, we have developed an effective yet gentle mechanical method compatible with 96-well plates using adaptive focused acoustics technology. This method was adapted for the release of Varicella-Zoster virus from MRC-5 cells and then applied to investigate infectious virus yield as a function of the cell density at infection. This microscale, high-throughput mechanical cell disruption method may be applicable to a variety of mammalian cell culture systems and intracellular products, thus expanding the scope of high-throughput screening.
authors.affiliation: Vaccine R&D, Merck & Co., Inc., West Point, PA 19486, USA.
authors.forename: May
authors.initials: M
authors.lastname: Ly
authors.affiliation: Vaccine R&D, Merck & Co., Inc., West Point, PA 19486, USA.
authors.forename: Franklin
authors.initials: F
authors.lastname: Lu
authors.affiliation: Vaccine R&D, Merck & Co., Inc., West Point, PA 19486, USA.
authors.forename: Gargi
authors.initials: G
authors.lastname: Maheshwari
authors.affiliation: Vaccine R&D, Merck & Co., Inc., West Point, PA 19486, USA. Electronic address: shyamsundar.subramanian@gmail.com.
authors.forename: Shyamsundar
authors.initials: S
authors.lastname: Subramanian
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line / Q000378:metabolism* / Q000821:virology; D003902:Detergents / Q000737:chemistry; D055808:Drug Discovery; D014645:Herpesvirus 3, Human / Q000254:growth & development; D057166:High-Throughput Screening Assays; D006801:Humans; D008322:Mammals
chemical_list: D003902:Detergents