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Updated: Apr 23, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Configuration, column construction, and column packing for a capillary liquid chromatography system
Abstract:
INTRODUCTIONCapillary or microcolumn (<0.5-mm internal diameter, I.D.) high-performance liquid chromatography (HPLC) is an extremely powerful technique for separating small quantities of proteins and peptides. This protocol describes a procedure for adapting conventional HPLC systems to provide accurate low-flow rates (0.4-4 μl/min) and gradients required to operate slurry-packed capillary columns. A key component of this system is a commercial axial-beam longitudinal flow cell that can be fitted to several commercial UV detectors. Procedures are described for the fabrication of 0.32-mm I.D. polyimide-coated fused-silica columns, slurry-packed with reversed-phase chromatographic supports.
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