“…The production process of the PEGylated lysozyme and the separation of the isoforms were carried out as recently described in detail by Blaschke et al…”
Section: Methodsmentioning
confidence: 99%
“…There is abundant literature on PEGylation chemistry, the influence of PEGylation on various properties of proteins and on the separation of proteins with different degrees of PEGylation . It has also been shown that hydrophobic interaction chromatography is a promising technique for the separation of PEGylated from unPEGylated proteins …”
“…The production process of the PEGylated lysozyme and the separation of the isoforms were carried out as recently described in detail by Blaschke et al…”
Section: Methodsmentioning
confidence: 99%
“…There is abundant literature on PEGylation chemistry, the influence of PEGylation on various properties of proteins and on the separation of proteins with different degrees of PEGylation . It has also been shown that hydrophobic interaction chromatography is a promising technique for the separation of PEGylated from unPEGylated proteins …”
“…Composition, structure, chemical surface, magnitude and signal of charges and protein hydration degree are important [8]. Calorimetric studies have shown that protein adsorption process on ion exchange sorbents is often endothermic, suggesting that other processes are taking place besides the expected ion-exchange interaction [36,[42][43][44]. So, it is important to obtain more in-depth knowledge on energy variations (enthalpy and entropy) involved in this process.…”
Section: Introductionmentioning
confidence: 98%
“…Many parameters, as pH, temperature, type and amount of salts, affects protein adsorption and only the knowledge of these effects allows a rational improvement of it [36]. Calorimetric techniques, mathematical correlations, computational modeling and others have been made in order to elucidate these mechanisms and their implications on the structure of proteins [8,36,[39][40][41][42].…”
“…Figure shows the results of the thermodynamic analysis of the data for di‐PEG‐lysozyme‐5kDa in a solution with 3500 mM sodium chloride, which is described closer in . The molar Gibbs energy of adsorption , which is calculated using the equilibrium data, is presented together with the associated molar enthalpy of adsorption , as measured in the calorimetric experiments, and the resulting molar entropy of adsorption .…”
Interaction of macromolecules in aqueous salt‐containing solution with a hydrophobic adsorbent is studied by adsorption equilibrium measurements and by independent isothermal titration calorimetry. The macromolecules are native as well as mono‐, di‐, and tri‐PEGylated lysozyme and four pure PEGs. The hydrophobic adsorbent is Toyopearl PPG‐600M. The salt is sodium chloride. The sodium chloride concentration in the aqueous 25 mM sodium phosphate buffer is varied from 2000 to 4500 mM at pH 7.0 and 25°C. PEGylation of the lysozyme is carried using 5 and 10 kDa PEG chains. The molar enthalpy of adsorption Δhp ads is calculated from the adsorption equilibrium and the calorimetric data. The results show that the adsorption of the PEGylated lysozyme is caused by both the interaction of the lysozyme and the interaction of the PEG chains with the adsorbent, respectively, but the interaction of the lysozyme is stronger than that of PEG. The comparison of the results of the present study on the influence of sodium chloride with a corresponding study on the influence of ammonium sulfate shows that the adsorption mechanism changes upon the variation of the salt. The knowledge of the adsorption mechanisms supports the systematic development of chromatographic purification steps.
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