title: Application of a new wall-less plate technology to complex multistep cell-based investigations using suspension cells.
abstract: Despite significant progresses, cell-based assays still have major limitations part to because of their plate format. Here, we present a wall-less plate technology based on unique liquid dynamics named DropArray that takes advantage of hydrophobic and hydrophilic surface properties. Liquid velocities within the DropArray plate were quantified through fluid dynamics simulation and complete retention of suspension cells experimentally demonstrated within the range of simulated shear stresses. Subsequently, we compared the DropArray technology with conventional microtiter plates in a cell-based protein-binding assay. Although the wall-less plate produced similar results with adherent cells, the advantage of the DropArray technology was absolutely clear when semiadherent or suspension cells were used in this multistep experimental procedure. The technology also was evaluated for the cell viability assay and generated similar results to conventional plate format while enabling significant reduction in toxic reagent use. Finally, we developed a DropArray cell-based assay to evaluate a bispecific antibody designed to engage cytotoxic T cells and trigger tumor cell killing. This assay enables for the first time the visualization and quantification of the specific killing events and represents a very powerful tool to further investigate functional aspects of the cancer immunotherapy.
authors.affiliation: Protein Chemistry Department, Genentech Inc, South San Francisco, CA 94080, USA.
authors.forename: Gabriel A
authors.initials: GA
authors.lastname: Quiñones
authors.forename: Travis I
authors.initials: TI
authors.lastname: Moore
authors.forename: Katrina
authors.initials: K
authors.lastname: Nicholes
authors.forename: Hyunjae
authors.initials: H
authors.lastname: Lee
authors.forename: Sudong
authors.initials: S
authors.lastname: Kim
authors.forename: Laura
authors.initials: L
authors.lastname: Sun
authors.forename: Noo Li
authors.initials: NL
authors.lastname: Jeon
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Stephan
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific; D001402:B-Lymphocytes / Q000276:immunology; D019556:COS Cells; D002460:Cell Line; D002470:Cell Survival; D002522:Chlorocebus aethiops; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D003601:Cytotoxicity Tests, Immunologic / Q000295:instrumentation / Q000379:methods; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007167:Immunotherapy; D020014:K562 Cells; D008213:Lymphocyte Activation; D009369:Neoplasms / Q000276:immunology / Q000628:therapy; D011485:Protein Binding; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D020298:U937 Cells
chemical_list: D018033:Antibodies, Bispecific