Surface energetics of protein adsorption onto chromatographic supports

  • Protein adsorption onto chromatographic supports has been studied utilizing the extended DLVO approach. Hydrophobic interaction chromatography was mainly explored due to it wide applicability in the recovery of a number of biologically significant macromolecules in its native form. The interactions between a number of selected model proteins and commercially available chromatographic beads i.e., Phenyl Sepharose 6 Fast Flow, Source 15 Phenyl, Toyopearl Phenyl 650-C and Toyopearl Butyl 650-C was studied via extended DLVO (XDLVO) calculations. The physicochemical properties of the proteins and the chromatographic beads, required for the calculations, are achieved by contact angle measurements and zeta potential determinations. Protein properties were investigated both at the hydrated and dehydrated state to achieve the colloidal-type calculation at the low and high salts. The mentioned approach allowed the calculations of interaction energy of proteins to chromatographic supports as a function of distance, at the operating buffer conditions. The XDLVO calculations were correlated with the actual separation behavior of the model proteins. The correlations plotted for all the chromatographic supports under study revealed that all the proteins can be segregated into two main groups; the proteins showing higher interaction energy minimum are eluting late in the chromatography while proteins showing low interaction energy are eluting earlier in the chromatographic experiments. Moreover, the calculations were able to expose the role of backbone chemistry harboring the same ligand and the effect of different ligands immobilized on the same supports. These studies have also shown the effect of the supporting surface used for protein immobilization. Thus, the XDLVO calculations were able to understand the underlying phenomenon and will suggest a broader tool to enhance a better understanding of the downstream bioprocessing, which will facilitate development, optimization and finally implementation of the bioprocess design.

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Publishing Institution:IRC-Library, Information Resource Center der Jacobs University Bremen
Granting Institution:Jacobs Univ.
Author:Muhammad Aasim
Referee:Marcelo Fernandez Lahore, J├╝rgen Fritz, Klaus Rischka
Advisor:Marcelo Fernandez Lahore
Persistent Identifier (URN):urn:nbn:de:101:1-201305294686
Document Type:PhD Thesis
Language:English
Date of Successful Oral Defense:2012/05/29
Year of Completion:2012
Date of First Publication:2012/06/22
PhD Degree:Biochemical Engineering
School:SES School of Engineering and Science
Library of Congress Classification:T Technology / TP Chemical technology / TP248.13-248.65 Biotechnology / TP248.3 Biochemical engineering. Bioprocess engineering
Call No:Thesis 2012/12

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