High performance flow injection analysis of recombinant protein G.
Chromatographic discs were investigated for their potential to substitute for the hitherto used cartridges in heterogeneous flow injection analysis. Originally designed for fast high performance liquid chromatography (HPLC) of biopolymers, the discs combine reliability with speed and resolution. This together with their price and their long-standing time made them attractive for use in flow injection analysis. The base material of the discs is a glycidyl methacrylate-co-ethylene dimethacrylate (GMA-EDMA) co-polymer. The epoxy groups inherent to this base structure can be used for immobilization purposes. In this first demonstration, antibodies were immobilized and the resulting affinity discs used for the fast analysis (< 5 min) of protein G from cell lysate of recombinant Escherichia coli. A linear calibration curve over several orders of magnitude as well as excellent reproducibility and correlation with data produced by conventional protein assay were obtained.
Hagedorn
J
J
Institute of Technical Chemistry, University of Hannover, Germany.
Kasper
C
C
Freitag
R
R
Tennikova
T
T
D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods; D007074:Immunoglobulin G / Q000032:analysis; D008024:Ligands; D008768:Methylmethacrylates; D009419:Nerve Tissue Proteins / Q000032:analysis*; D011108:Polymers; D011994:Recombinant Proteins / Q000032:analysis
C029152:G-substrate; D007074:Immunoglobulin G; D008024:Ligands; D008768:Methylmethacrylates; D009419:Nerve Tissue Proteins; D011108:Polymers; D011994:Recombinant Proteins; C020347:GMA-EDMA