Development and optimization of a process for automated recovery of single cells identified by microengraving |
| |
Authors: | Jae Hyeok Choi Adebola O Ogunniyi Mindy Du Minna Du Marcel Kretschmann Jens Eberhardt J Christopher Love |
| |
Institution: | 1. Dept. of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;2. Dept. of Biology, Whitehead Institute for Biomedical Research, 9 Cambridge Center, Massachusetts Institute of Technology, Cambridge, MA 02139;3. AVISO GmbH, Stockholmer Stra?e 10, D‐07747 Jena, Germany |
| |
Abstract: | Microfabricated devices are useful tools for manipulating and interrogating large numbers of single cells in a rapid and cost‐effective manner, but connecting these systems to the existing platforms used in routine high‐throughput screening of libraries of cells remains challenging. Methods to sort individual cells of interest from custom microscale devices to standardized culture dishes in an efficient and automated manner without affecting the viability of the cells are critical. Combining a commercially available instrument for colony picking (CellCelector, AVISO GmbH) and a customized software module, we have established an optimized process for the automated retrieval of individual antibody‐producing cells, secreting desirable antibodies, from dense arrays of subnanoliter containers. The selection of cells for retrieval is guided by data obtained from a high‐throughput, single‐cell screening method called microengraving. Using this system, 100 clones from a mixed population of two cell lines secreting different antibodies (12CA5 and HYB099‐01) were sorted with 100% accuracy (50 clones of each) in ~2 h, and the cells retained viability. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 |
| |
Keywords: | automation high‐throughput screening clonal selection monoclonal antibody |
|
|