High performance flow injection analysis of recombinant protein G

Chromatographic discs were investigated for their potential to substitute f or the hitherto used cartridges in heterogeneous Bow injection analysis. Or iginally 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 attractiv e for use in flow injection analysis, The base material of the discs is a g lycidyl 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 cur ve over several orders of magnitude as well as excellent reproducibility an d correlation with data produced by conventional protein assay were obtaine d. (C) 1999 Elsevier Science B.V. All rights reserved.