TSK-GEL

SIZE EXCLUSION CHROMATOGRAPHY USING TSK-GEL COLUMNS AND TOYOPEARL RESINS... 

I. SILICA-BASED TSK-GEL SW/SWxl COLUMNS FOR GEL-FILTRATION CHROMATOGRAPHY (GFC)... 

TABLE 4.1 Properties and Separation Ranges for TSK-GEL SW Type Packings"... 

The different pore sizes and exclusion limits of each TSK-GEL SW column will have a substantial effect on the resolution of a biomolecule mixture. G2000SW packing, which has the smallest pores, provides the best resolution for smaller proteins such as myoglobin and cytochrome c (16,900 and 12,400 Da, respectively, Rs = 1.01). Resolution of the proteins myoglobin... 

FIGURE 4.2 Polyethylene oxide, dextran, and protein calibration curves for TSK-GEL SW Columns. Column TSK-GEL SW, two 7.S mm x 60 cm columns in series. Sample , proteins Q, polyethylene oxides O, dextrans. Elution dextrans and polyethylene oxides distilled water proteins 0.3 A1 NaCI in 0.1 M phosphate buffer, ph 7. Flow rate 1.0 ml/min. Detection UV at 220 nm and Rl. 

TABLE 4.2 Comparison of Protein Resolution Factors (Rs) on TSK-GEL SW Columns"... 

TABLE 4.4 Effect of Buffer Composition on Mass Recovery of Proteins from TSK-GEL G3000SW< ... 

FIGURE 4.4 pH stability of TSK-GEL SW packings. Conditions I g of each TSK-GEL SW packing was suspended in 10 ml of buffer at indicated pH for I month. The concentration of silica dissolved in each buffer was then determined. 

Elow rate determines the separation time and can significantly affect resolution and efficiency. The effect of flow rate on HETP for TSK-GEL SW and TSK-GEL SWxi analytical columns is shown in Fig. 4.6. Resolution is typically higher at slower flow rates, although results shown in Fig. 5B indicate that, with increasing sample load, the faster flow rates can give higher resolution. 

FIGURE 4.7 Separation of mixture of proteins and peptides on a TSK-GEL G2000SWxl column using Wyatt/Optilab DSP and miniDawn detectors. 

TSK-GEL SW columns have also been used to isolate heavy and light chains from immunoglobulins. Kast et al. (7) reported how these chains were desalted and isolated on a 21.5-mm i.d. X 60-cm TSK-GEL G4000SW semipreparative column using a 0.1 trifluoroacetic acid (TEA)/40% acetonitrile mobile phase. 

Also available are semipreparative, 21.5-mm i.d. and preparative 55-, 108-, 158-, and 210-mm i.d. stainless-steel columns packed with TSK-GEL SW parti-... 

TSK-GEL PW type columns are commonly used for the separation of synthetic water-soluble polymers because they exhibit a much larger separation range, better linearity of calibration curves, and much lower adsorption effects than TSK-GEL SW columns (10). While TSK-GEL SW columns are suitable for separating monodisperse biopolymers, such as proteins, TSK-GEL PW columns are recommended for separating polydisperse compounds, such as polysaccharides and synthetic polymers. 

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