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Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays
Authors:Bannister David  Wilson Andrew  Prowse Leah  Walsh Meg  Holgate Rob  Jermutus Lutz  Wilkinson Trevor
Affiliation:Cambridge Antibody Technology, Granta Park, Milstein Building, Cambridge CB1 6GH, UK. david.bannister@cambridgeantibody.com
Abstract:We describe a method for high-throughput, parallel purification of secreted proteins to analyse large numbers of protein samples in cell-based assays for the discovery of protein therapeutics. The procedure is generic and capable of 96 parallel purifications and compatible, in both yield and purity, with a wide assay range. By optimising expression and purification steps as well as using novel hardware, in particular a chromatography press capable to purify target proteins from viscous media, we exemplify the process for the generation of single-chain Fv antibody fragments (scFv) and the purification of full-length IgG. The described process can operate robustly with a throughput of over 2,000 samples per month.
Keywords:scFv  purification  IgG  His Tag  phage display  IMAC  cation exchange chromatography
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