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

David Bannister, Andrew W. Wilson, Leah Prowse, Meg Walsh, Rob Holgate, Lutz Jermutus, Trevor Wilkinson

Biotechnology and Bioengineering · 2006 · 18 citations · 10 references

Concepts

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.

References

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