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Extended DLVO calculations expose the role of the structural nature of the adsorbent beads during chromatography
Author(s) -
Aasim Muhammad,
Bibi Noor Shad,
Vennapusa Rami Reddy,
FernandezLahore Marcelo
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100719
Subject(s) - dlvo theory , chemistry , chromatography , adsorption , protein adsorption , lysozyme , electrochromatography , organic chemistry , biochemistry , colloid , capillary electrophoresis , capillary electrochromatography
Protein adsorption onto hydrophobic interaction chromatography supports was studied by a surface‐thermodynamics approach. To gather relevant experimental information, contact angle measurements and zeta potential determinations were performed on three different commercial adsorbent beads, P henyl S epharose 6 F ast F low, T oyopearl P henyl 650‐C and S ource 15 P henyl, having soft to rigid backbone structure. Similar information was obtained for a collection of model proteins, lysozyme, bovine serum albumin ( BSA ), polygalacturonase, aminopeptidase, chymosin, aspartic protease, beta‐galactosidase, human immunoglobulin G , and lactoferrin, were evaluated in the hydrated and in the dehydrated state. Based on the mentioned experimental data, calculations were performed to obtain the (interfacial) energy versus distance profiles of nine individual (model) proteins on (commercial) beads of three different types. All of these beads harbored the phenyl‐ligand onto a matrix of differentiated chemical nature. Extended D erjaguin, L andau, V erwey, and O verbeek ( DLVO ) calculations were correlated with actual chromatographic behavior. Typical chromatography conditions were employed. The population of model proteins utilized in this study could be segregated into two groups, according to the minimum values observed for the resulting interaction energy pockets and the corresponding retention volumes (or times) during chromatography. Moreover, trends were also identified as a function of the type of adsorbent bead under consideration. This has revealed the influence of the physicochemical nature of the bead structure on the adsorption process and consequently, on the expected separation behavior.