Calibration of Size-Exclusion Chromatography: Use of a Double Gaussian Distribution Function to Describe Pore Sizes

✍ Scribed by J.E. Harlan; D. Picot; P.J. Loll; R.M. Garavito

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 589 KB
• Volume: 224
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1995.1087

✦ Synopsis

A method for the calibration of size-exclusion chromatographic columns is proposed that takes into account the nonlinear dependence of the Stokes radius (R_{\text {* }}) upon the partition coefficient (K_{\mathrm{p}}). The method is based on the assumption that the pore size distribution of a molecular sieve column can be described by the sum of two Gaussian distributions and has been successfully tested with low-pressure chromatography gels (Sephacryl and Superose) and HPLC gels (TSK SW) over a wide range of protein sizes. An application of this method is described, in which aggregation states of the membrane protein prostaglandin (\mathrm{H}_{2}) synthase solubilized in nonionic detergents are estimated. (\odot 1985) Academic Pross, Inc.