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Updated: Jun 2, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
A high throughput approach to purifying chiral molecules using 3μm analytical chiral stationary phases via
Chris Hamman1, Mengling Wong, Michael Hayes
1Discovery Chemistry, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA. hamman.christopher@gene.com
Abstract:
In this study we describe a fast 2.5 min gradient chiral screening method that utilizes 3 μm particles CSPs. An empirical approach to scale-up from the 2.5 min gradient method to an isocratic preparative method is described. We also evaluate the use of 5 μm preparative columns that are 150 mm in length versus the industry standard of 250 mm. Finally, we evaluate eleven different CSPs against 46 compounds, 23 commercially available and 23 internal compounds from a variety of projects. All 46 compounds were separated using the 2.5 min gradient method. Assuming an R(s) of 1.0 or greater, the Chiralpak AD column from Chiral Technologies proved most useful, followed by the Cellulose-1 from Phenomenex. The Cellulose-4, a novel stationary phase from Phenomenex, provided the third most separations of the eleven columns tested. For the 46 compounds tested, the Chiralcel OJ column from Chiral Technologies outperformed Phenomenex's version, the Lux Cellulose-3.
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