Cyto-Mine: An Integrated, Picodroplet System for High-Throughput Single-Cell Analysis, Sorting, Dispensing, and Monoclonality Assurance
Dimitris N. Josephides, Serena Davoli, W. T. Whitley, Raphael Ruis, Robert B. Salter, Sinan Gokkaya, M Vallet, Darren Matthews, Giuseppe Benazzi, Elena Shvets, Frank Gesellchen, Drew Geere, Xin Liu, Xin Li, Benedict Mackworth, William Young, Zachary Owen, Clive A. Smith, Dale Starkie, James H. White, Bernie Sweeney, Matthew Hinchliffe, Simon Tickle, Daniel Lightwood, M. Řehák, Frank F. Craig, David Holmes
Abstract
The primary goal of bioprocess cell line development is to obtain high product yields from robustly growing and well-defined clonal cell lines in timelines measured in weeks rather than months. Likewise, high-throughput screening of B cells and hybridomas is required for most cell line engineering workflows. A substantial bottleneck in these processes is detecting and isolating rare clonal cells with the required characteristics. Traditionally, this was achieved by the resource-intensive method of limiting dilution cloning, and more recently aided by semiautomated technologies such as cell sorting (e.g., fluorescence-activated cell sorting) and colony picking. In this paper we report on our novel Cyto-Mine Single Cell Analysis and Monoclonality Assurance System, which overcomes the limitations of current technologies by screening hundreds of thousands of individual cells for secreted target proteins, and then isolating and dispensing the highest producers into microtiter plate wells (MTP). The Cyto-Mine system performs this workflow using a fully integrated, microfluidic Cyto-Cartridge. Critically, all reagents and Cyto-Cartridges used are animal component-free (ACF) and sterile, thus allowing fast, robust, and safe isolation of desired cells.