Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays.
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.
Cambridge Antibody Technology, Granta Park, Milstein Building, Cambridge CB1 6GH, UK. david.bannister@cambridgeantibody.com
David
D
Bannister
Andrew
A
Wilson
Leah
L
Prowse
Meg
M
Walsh
Rob
R
Holgate
Lutz
L
Jermutus
Trevor
T
Wilkinson
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001681:Biological Assay / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D011994:Recombinant Proteins / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins