Protein surface area and retention in hydrophobic interaction chromatography

Katti, Anita M.; Maa, Yih-Fen; Horváth, Cs.

doi:10.1007/bf02688560

Cited by 39 publications

(19 citation statements)

References 22 publications

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“…The theory predicts that, at sufficiently high salt concentrations, where the mechanism of retention is governed predominantly by hydrophobic interactions, the retention increases with both the molal salt concentration in the eluent and the size of the eluite or its hydrophobic moiety. Experimental data (52,64,66,(69)(70)(71) for protein retention obtained with "antichaotropic", also called "cosmotropic", salts (72), which do not preferentially bind to the protein, lend support to the key features of the theory.…”

Section: Hydrophobic Interaction Chromatographymentioning

confidence: 52%

Retention Thermodynamics in Hydrophobic Interaction Chromatography

and

Horváth

1996

Ind. Eng. Chem. Res.

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This paper is a review of the thermodynamics of retention in hydrophobic interaction chromatography (HIC) with mildly hydrophobic stationary phases and aqueous salt solutions usually employed in protein purification. Since the role of salt in HIC has been well documented, our focus was to investigate the temperature effect on the retention behavior in HIC and to compare the results with those obtained for other processes driven by the hydrophobic effect. Using nonpolar dansyl amino acids as model compounds, retention data obtained on three stationary phases yielded nonlinear van't Hoff plots in the temperature range from 5 to 50°C. Thermodynamic analysis of the data revealed significant heat capacity effects. The enthalpy and entropy changes were large and positive at low temperatures, decreased with increasing temperature, and became negative at high temperatures. The results parallel those of calorimetric studies on other processes based on the hydrophobic effect, such as dissolution in water of nonpolar liquids, gases, and solids as well as protein folding. Thermodynamic parameters from HIC measurements also confirmed the existence of certain exothermodynamic relationships, such as enthalpy-entropy compensation and molecular area correlations. In order to examine at the molecular level the energetics of HIC retention as well as the dissolution of nonpolar gases in water, the pertinent thermodynamic parameters were expressed in terms of nonpolar molecular area and interfacial tensions by employing the solvophobic theory. It was found that these expressions from HIC and dissolution data are nearly identical, thus confirming the mechanistic identity of the two processes.

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Section: Hydrophobic Interaction Chromatographymentioning

confidence: 52%

Retention Thermodynamics in Hydrophobic Interaction Chromatography

and

Horváth

1996

Ind. Eng. Chem. Res.

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“…Hence, the EC-SODs of Scots pine are characterized by additional basic groups compared with symplasmic SODs. The EC-SODs showed lower surface hydrophobicity than chloroplastic and cytosolic SODs of Scots pine, as indicated by hydrophobic interaction chromatography (Katti et al 1987).…”

Section: Discussionmentioning

confidence: 95%

Pinus sylvestris L. needles contain extracellular CuZn superoxide dismutase

Streller

Wingsle

1994

Planta

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Four new isoforms of superoxide dismutase (SOD; superoxide: superoxide oxidoreductase, EC 1.15.1.1.) were identified in extracellular washing fluid from Scots pine (Pinus sylvestris L.) needles. The isoforms had an apparent molecular mass of 33 kDa. No neutral carbohydrates were present in the enzymes. The enzymatic activities were inhibited by 3 mM NaCN. One of the putative extracellular SOD isoforms was purified and NH2-terminal-sequenced. The sequence contained the domain KAVAVL. The domains VEG and V(K/S)G, present in chloroplastic and cytosolic CuZn SODs of plants, respectively, were not detected. The enzyme was composed of two subunits of 17.8 kDa each. The isoelectric point was determined to be 6.5. The results suggest the existence of an extracellular SOD in Scots pine.

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“…While this may actually be an advantage in some applications, we have recently obviated this need by adapting PEG-ABS to a solid-supported chromatographic mode (23,24). Although the chromatographic techniques employed probably fit under the category of hydrophobic interaction chromatography (25,26), we prefer to name this separation technique: Aqueous Biphasic Extraction chromatography (ABEC). We believe this acronym is appropriate due to the behavior of our resins and the strong relationship with liquidfliquid PEG-ABS.…”

Section: Introductionmentioning

confidence: 99%

New Technetium-99M Generator Technologies Utilizing Polyethylene Glycol-Based Aqueous Biphasic Systems

Rogers

Bond

Zhang

et al. 1997

Separation Science and Technology

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Two new schemes for TcOJMo0~-separations fiom OH-and MOO: -media using polyethylene glycol (PEG)-based aqueous biphasic systems (ASS) have been developed. The two most important salt solutions in current -Tc-generator technologies, OH-and MOO: , also salt out PEG to form ABS. In liquidfliquid PEG-ABS, pertechnetate can be separated from molybdate with separation factors as high as 10,000. Stripping is accomplished by reduction of the TcO; and back extraction into a salt solution. The strip solution can be the salt of an imaging agent (e.g., Na,HEDPA) and thus may, under the appropriate conditions, be injected directly into the human body. -TcOi can also be concentrated fiom a dilute load solution of ?MOO: in NaOH using an aqueous biphasic extraction chromatographic technique (ABEC). A rinse with KJO, assures that all ?MOO:-is removed from the column and this is confirmed by a rapid drop in ?Mo activity by the fourth free column volume (fcv) of rinse. The T c O i is then eluted with water. This chromatographic separation affords 94% of the V c O ; activity in 5 fcv, with the y spectrum showing less than 2 x IO4 of the original %o activity.

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Protein surface area and retention in hydrophobic interaction chromatography

Cited by 39 publications

References 22 publications

Retention Thermodynamics in Hydrophobic Interaction Chromatography

Retention Thermodynamics in Hydrophobic Interaction Chromatography

Pinus sylvestris L. needles contain extracellular CuZn superoxide dismutase

New Technetium-99M Generator Technologies Utilizing Polyethylene Glycol-Based Aqueous Biphasic Systems

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