Screening for peptide affinity ligands on CIM monoliths.
Screening of peptide ligands for affinity chromatography usually involves incubation with the target protein in a batch system. In an additional step, peptides with fast binding kinetics have to be selected in respect to satisfactory performance under flow conditions on a support ensuring optimal three-dimensional presentation of the peptide. We have developed a rapid screening system based on peptide synthesis and screening on CIM((R)) disks. The disk size was minimized to fit into microplates usually applied for solid-phase extraction. In combination with a vacuum manifold, semi-automated peptide synthesis and screening for binding to a target protein under simulated chromatography conditions are possible. Various analytical methods can be applied for parallel and automated determination of the quantity, integrity, or activity of the target protein in the flow through or bound to the affinity support. This system also allows parallel screening for suitable chromatographic conditions like running buffer, washing, and elution conditions.
Pflegerl
K
K
Institute of Applied Microbiology, University of Agricultural Sciences, Muthgasse 18, Vienna, Austria.
Podgornik
A
A
Berger
E
E
Jungbauer
A
A
D002352:Carrier Proteins; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D004800:Enzymes, Immobilized; D005169:Factor VIII / Q000032:analysis*; D008024:Ligands; D008768:Methylmethacrylates; D010455:Peptides / Q000032:analysis* / Q000138:chemical synthesis*; D019809:Streptavidin / Q000032:analysis* / Q000235:genetics
D002352:Carrier Proteins; D004800:Enzymes, Immobilized; D008024:Ligands; D008768:Methylmethacrylates; D010455:Peptides; C020347:GMA-EDMA; D005169:Factor VIII; D019809:Streptavidin