Optimum support properties for protein separations by high-performance size exclusion chromatography.

摘要: Abstract Optimum chromatographic properties of high performance size exclusion chromatography (HPSEC) proteins, such as resolution, molecular weight accuracy and recovery, are obtained on packings columns with tailor-made physical chemical structures, employed at properly adjusted eluent compositions operation conditions. SEC-theory suggests that a broad fractionation range linearity the log—linear calibration plot can be achieved by use two (10- 80-nm pore size, characterized pore-size distribution (psd) equal to or less than 1 decade internal column porosity ϵ p ), rather single 30- 50-nm packing wide psd. Favourably high-phase ratios /ϵ o ⪢/ 1.0 for HPSEC were accomplished minimum interstitial value (the specific volume, ν , multiplied density, ϱ .) Ligands diol, N -acetoxyamino oligomeric ether propyl- propoxy-spacer bonded silica highest density appear provide mass recovery bioactivity well stability. Such packings, available in 3–5 μm particle ranges narrow distribution, packed into ⪢6 mm i.d. ⪢500 length, offer best compromise respect speed pressure drop. More careful studies required explain effects protein conformational changes interconversions during elution columns.