Interplay of hydrophobic and electrostatic interactions in biopolymer chromatography

Melander, Wayne R.; Rassi, Ziad El; Horváth, Csaba

doi:10.1016/s0021-9673(01)96437-4

Cited by 250 publications

(117 citation statements)

References 25 publications

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“…The solvophobic theory is based on the association and solvation of the participating species and assumes that the molal surface tension increment of the salt determines solute retention (Melander and Horvath, 1977;Melander et al, 1984Melander et al, , 1989. In the solvophobic theory, it is believed that the cavity is formed and then closed on the stationary and mobile phases, which is not always valid if the salt has a strong interaction with the proteins.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Xia

Nagrath

Garde

et al. 2004

Biotech & Bioengineering

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It is well established that salt enhances the interaction between solutes (e.g., proteins, displacers) and the weak hydrophobic ligands in hydrophobic interaction chromatography (HIC) and that various salts (e.g., kosmotropes, chaotropes, and neutral) have different effects on protein retention. In this article, the solute affinity in kosmotropic, chaotropic, and neutral mobile phases are compared and the selectivity of solutes in the presence of these salts is examined. Since solute binding in HIC systems is driven by the release of water molecules, the total number of released water molecules in the presence of various types of salts was calculated using the preferential interaction theory. Chromatographic retention times and selectivity reversals of both proteins and displacers were found to be consistent with the total number of released water molecules. Finally, the solute surface hydrophobicity was also found to have a significant effect on its retention in HIC systems. B 2004 Wiley Periodicals, Inc.

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Section: Introductionmentioning

confidence: 99%

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Xia

Nagrath

Garde

et al. 2004

Biotech & Bioengineering

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“…Protein retention in HIC has been interpreted in the light of the underlying thermodynamic phenomena, by considering the effect of salt. Melander et al (1989) Melander et al (1989) to describe the effect of salt concentration on macromolecule retention in chromatography (IEC and HIC) can be applied to any stationary phase consisting of a highly hydrated surface modified with charged ligands (in the case of IEC), weakly hydrophobic moieties (in the case of HIC), or both. Electrostatic and hydrophobic interactions between the macromolecule and the stationary phase are treated separately.…”

Section: Retention Mechanisms In Hydrophobic Interaction Chromatographymentioning

confidence: 99%

Hydrophobic Interaction Chromatography: Fundamentals and Applications in Biomedical Engineering

Mahn¹

2012

Biomedical Science, Engineering and Technology

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“…Thus, the chromatographic behavior is not necessarily governed by an overall property of the protein, such as pI. There are also other phenomena occurring in the chromatofocusing technique that contribute to the retention mechanism, including Donnan potential [34,35], hydrophobic interaction [36], and buffer concentration [25,26]. It should be noted that the buffer concentration can greatly enhance the separating capability of the technique.…”

Section: Chromatofocusing Aspectsmentioning

confidence: 99%

Gradient chromatofocusing-mass spectrometry: A new technique in protein analysis

Lian

Hribar

Zhou

et al. 2008

J. Am. Soc. Mass Spectrom.

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A new analytical technique, gradient chromatofocusing-mass spectrometry (gCF-MS), was developed employing ion-exchange high-performance liquid chromatography (HPLC) interfaced to an electrospray-quadrupole mass spectrometer in the determination of proteins. There have been few reports, if any, of a HPLC-MS technique for proteins in which the ion-exchange column is directly interfaced to the mass spectrometer. The employment of a linear pH gradient elution scheme directly interfaced to mass spectrometry is also unique in the present work. The technique was demonstrated by the separation of six proteins (carbonic anhydrase II, enolase, ␤-lactoglobulin A, lactoglobulin B, soybean trypsin inhibitor, and amyloglucosidase) employing a descending linear pH gradient from pH 9 to 2.6 on a 50 mm ϫ 2.1 mm DEAE HPLC column using volatile buffer components. A signal enhancement solution consisting of 8% formic acid in acetonitrile was pumped post-column and was mixed 1:1 with column effluent and then directed on-line into the mass spectrometer. Molecular masses of the proteins were determined within Ϯ0.010% to 0.033% (Ϯ100 to 330 ppm) with peak height total ion current detection limits of 4 to 78 pmol of injected amounts (S/N ϭ 3). This technique is applicable to the analysis of proteins and other charged molecules. T here is great interest in developing a mass spectrometry (MS)-compatible high-performance liquid chromatography (HPLC) technology which performs charge-based separations, a principal dimension in the 2D-gel electrophoresis technique, for application in proteomic and protein characterization studies. Development of such a HPLC technique would be progress towards addressing the limitations of the 2D-gel electrophoresis technique, which has been hampered by its design incompatibility for direct interfacing with MS (requiring excision of each protein band from the gel), limitation in quantitative dynamic range, inability of determining small proteins (5-8000 Da), high labor intensity, long sample run times, as well as other disadvantages [1,2].Development of charged-based HPLC techniques directly coupled to mass spectrometry (MS) has been a difficult challenge. This is because the mobile phase salts commonly used in ion-exchange chromatography, the most used charge-based HPLC technique, suppress the MS signal. The most utilized approach incorporating ion-exchange chromatography with mass spectrometry has been to use a 2D ion-exchange/reversed-phase HPLC technique, directing the ion-exchange fractions to a reversed-phase column for further separation and removal of salt before on-line mass spectrometry. This has been applied to proteomic studies in the analysis of proteins [3] and protein digests [4,5]. A "stair step" gradient is employed for the ion-exchange dimension. This first dimension thus serves as a crude initial separation step, and is not the stage that is directly interfaced to the mass spectrometer.The application of two-dimensional chromatography using the combination of chromatofocusing and reversed-p...

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Interplay of hydrophobic and electrostatic interactions in biopolymer chromatography

Cited by 250 publications

References 25 publications

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Hydrophobic Interaction Chromatography: Fundamentals and Applications in Biomedical Engineering

Gradient chromatofocusing-mass spectrometry: A new technique in protein analysis

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