Hydrophobic Chromatography

Hydrophobic Interaction Chromatography

“…The method of Shaltiel (1974), in which a gel was selected from a homologous chain length series and then operated at low ionic strength with elution by deforming, that is, denaturing buffers was soon abandoned in favour of the method of Porath et al (1973) and Hjerten (1973), who proposed column operation at high salt concentrations and elution by a negative salt gradient (see Hjerten et al (1974) and Rosengren et al (1975)). This method is still widely employed today.…”

“…Here, the preparation of a quantised homologous hydrophobicity gradient in the form of a library for chromatographic purposes will be described. As stated above, there are two methods (Figure 1b and c) for the synthesis of controlled hydrophobicity gels: (a) via homologous series procedure (variation of alkyl chain length (seen, Shaltiel (1974Shaltiel ( , 1984)) and (b) the surface concentration series procedure (variation of the alkyl surface concentration, see Jennissen and Heilmeyer (1975) and Jennissen (2000b)). …”

“…Er-El et al (1972) and Yon (1972) independently reported the chromatographic separation of proteins by way of hydrophobic interactions. In both the cases the hydrophobic matrix consisted of agarose (Sepharose 4B, Pharmacia, now GE-Healthcare) to which amino alkane derivatives had been coupled by the cyanogen bromide (CNBr) method (for reviews, see (Shaltiel, 1984;Yon, 1977;Jennissen, 1995)). The surprising result in Shaltiel's experiments (Er-El et al, 1972) was that a very normal hydrophilic enzyme, phosphorylase b, could be purified on a C 4 -hydrocarbon-coated agarose to near homogeneity in one step, implicitly questioning the general doctrine of the time that all hydrophobic amino acids are buried in the interior of proteins.…”

Thermodynamic analysis of the interaction between proteins and solid surfaces: application to liquid chromatography

“…The theoretical discussion presented below has its origins in the substantial body of scientific contributions from various investigators 23,24,28,[32][33][34][35]38,39,[66][67][68][80][81][82][83][84][85][86][87] interested in the relationship between the intra-and inter-facial behaviour of proteins or polypeptides, their interactions with solvents and ionic species in the bulk solution phase and with chemically defined media, such as chromatographic sorbents, and their underpinning thermodynamic and biophysical dependencies. Particularly noteworthy contributions concerning the role of hydrophobic interactions in HIC and RPC chromatographic processes with proteins and polypeptides have been made by Shaltiel et al, 88,89 Jennissen et al, 90,91 Hearn et al [33][34][35][36] and Horvath and coworkers. 23,24,28,38,39,66,80,82 Figure 3 illustrates a comprehensive framework for interpreting and interlinking thermodynamic and extra-thermodynamic data derived from liquid chromatographic measurements with polar/non-polar analytes, such as amino acids, peptides and proteins in HIC and RPC systems.…”

Affinity Chromatography: Principles and Applications

“…Some say it would be more accurate if termed bioaffinity chromatography (O'Carra et al, 1974) or hydrophobic affinity (Shaltiel, 1974). Nonetheless, the term affinity chromatography has been expanded to describe a potential method of separating biomolecule mixtures on the basis of specific biological interactions.…”