Salting-out in Amphiphilic Gels as a New Approach to Hydrophobia Adsorption

Porath, Jerker; Sundberg, Lars; Fornstedt, Nermin; Olsson, Ingmar

doi:10.1038/245465a0

Cited by 218 publications

(61 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Significant changes in peak position due to increasing salt concentration have been reported [32,33]. In these reports, the properties or the adsorbent used were different from ours.…”

Section: Effect Of S I L Tcontrasting

confidence: 50%

Non‐ionic Adsorptive Immobilization of Proteins to Palmityl‐Substituted Sepharose 4B

Nemat‐Gorgani

Karimian

1982

European Journal of Biochemistry

View full text Add to dashboard Cite

1. Palmityl-substituted Sepharose 4B prepared by the glycidyl ether method of S. Hjerten et al. [ J . Chromutogr. (1974) 101, 281 -2881 has been used as a non-ionic matrix for protein adsorption. A number of proteins, some of which are catalytically active, may be immobilized on this adsorbent in the form of suspension or column.2. Of the proteins examined, bovine serum albumin, hemoglobin, myoglobin, lysozyme, glutamate dehydrogenase, and P-galactosidase were immobilized on the adsorbent irrespective of NaCl concentration. Trypsin, a-chymotrypsin, papain, pepsin, and amyloglucosidase were totally adsorbed either in the absence of any additional salt or at high salt concentrations. Cytochrome c, used as a model protein, was totally immobilized only at high ionic strength and low pH.3. Immobilization normally took place with an apparent increase in initial activity rates. In the case of trypsin using N "-benzoyl-DL-arginine p-nitroanilide as substrate, adsorption resulted in an increase in V(app).4. Beef-liver glutamate dehydrogenase, used as a model allosteric enzyme, was found to retain its allosteric properties towards ADP and GTP after immobilization.5. Results are discussed in terms of specific interactions involving a smaller number of binding sites in protein molecules as compared to the multiple attachment to highly substituted adsorbents prepared with shorter ligands. Retention of the essential properties of the proteins examined in this study are ascribed to these characteristics of the adsorbent and to its non-ionic nature. Relevance of these observations to in vivo processes and the potential use of the adsorbent for enzyme immobilization are also discussed.In recent years, much attention has been directed towards use of hydrophobic properties of proteins for their purification. In most cases, the proteins of interest are bound at high salt concentrations to agarose derivatives containing hydrophobic ligands, and are eluted at lower salt concentrations. Adsorption and desorption thus correspond to the general 'salting out' and 'salting in' properties of proteins [I]. In this so-called 'hydrophobic interaction chromatography' the degree of interaction depends not only on the ionic strength, but also on the relative hydrophobicities of the protein and the ligand, the nature of the salt in the medium, the ligand concentration, and the ambient temperature.

show abstract

“…Significant changes in peak position due to increasing salt concentration have been reported [32,33]. In these reports, the properties or the adsorbent used were different from ours.…”

Section: Effect Of S I L Tcontrasting

confidence: 50%

Non‐ionic Adsorptive Immobilization of Proteins to Palmityl‐Substituted Sepharose 4B

Nemat‐Gorgani

Karimian

1982

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…But we observed that the control gel to which only mercaptoethanol has been coupled showed very distmct group adsorption properties toward specific serum protems, m partrcular the rmmunogloblns and ~~-macroglobulIns Further mvesttgatlons revealed that the adsorptron obtamed IS salt dependent and mcreases stgmftcantly m the presence of high concentrattons of neutral salts This saltmg-out chromatography 1s remmrscent of one of the condttlons set forth for hydrophobic mteractlon chromatography [3] and, therefore, a comparative study usmg a commercially available hydrophobic adsorbent was made. Our results mdrcate that the adsorptton charactertsttcs of the new adsorbent are dtstmct from those based on hydrophobtc Interaction.…”

Section: Introductionmentioning

confidence: 99%

Thiophilic adsorption ‐ a new method for protein fractionation

1985

Self Cite

View full text Add to dashboard Cite

Dlvlnyisulphone-activated agarose to which mercaptoethanol IS coupled showed very setectlve group adsorption of human serum protems, m particular the ~mmunogIobul~ns The adsorptxon Increases markedly m the presence of high concentrations of neutral water-structure formmg salts and IS dlstmct from adsorptions based on hydrophobic mteractlon A characterlstlc feature of this new type of adsorbent 1s the structure of the groups attached to the polymer, @, I e , R-S-CH,-CH,-SO,-CH,-cH,-O-Q, where R IS a small ahphatlc residue Our results mdlcate that the thloether sulphur and the adJacent sulphone group act cooperatlvely and are apparently necessary to mamtam the dlstmct behavlour of such dbsorbents

show abstract

“….). Sepharose columns carrying hydrophobic groups have been examined for this purpose ('hydrophobic chromatography' [ 11, 'phosphate induced protein chromatography' [2], 'hydrophobic salting-out chromatography' [3]). Unfortunately, this method has been successfully applied to a too small number of proteins to be considered as general.…”

mentioning

confidence: 99%