Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays

Biotechnol Bioeng. 2006 Aug 5;94(5):931-7. doi: 10.1002/bit.20914.

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.

Publication types

  • Evaluation Study

MeSH terms

  • Antibodies, Monoclonal / isolation & purification*
  • Biological Assay / methods*
  • Chromatography, High Pressure Liquid / instrumentation*
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, Ion Exchange / instrumentation*
  • Chromatography, Ion Exchange / methods
  • Equipment Design
  • Equipment Failure Analysis
  • Microfluidics / instrumentation*
  • Microfluidics / methods
  • Recombinant Proteins / isolation & purification

Substances

  • Antibodies, Monoclonal
  • Recombinant Proteins