Microcalorimetric study of adsorption of human monoclonal antibodies on cation exchange chromatographic materials

Dieterle, Michael; Blaschke, Tim; Hasse, Hans

doi:10.1016/j.chroma.2008.07.023

Cited by 30 publications

(32 citation statements)

References 31 publications

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“…The microcalorimetric measurements were carried out with an ITC4200 microcalorimeter (Calorimetry Science Corp., Lindon, NY, USA) as previously described in [9]. The 1 ml sample ampoule was filled with the desired amount of adsorbent (20-80 L) using a MediaScout ResiQuot device (Atoll GmbH, Weingarten, Germany).…”

Section: Isothermal Titration Calorimetrymentioning

confidence: 99%

“…The evaluation of the ITC data and the calculation of the specific enthalpy of adsorption were carried out as previously described in [9] and are not discussed here in detail. Preliminary experiments showed that the contribution of the adsorption of buffer ions and the heat of dilution of buffer ions to the measured signal can be neglected here.…”

Section: Evaluation Of Itc Experimentsmentioning

confidence: 99%

“…Much work has been done in the past to overcome this drawback, especially calorimetric techniques were shown to yield results which contribute to a better understanding of the adsorption mechanisms. Dieterle et al [9] have shown that the adsorption of monoclonal antibodies on cation exchange resins is far from being a strongly exothermal process driven by ionic forces. Similar results were found in other studies on the adsorption energetics of proteins on chromatographic resins especially at high salt concentrations, adsorber loadings and near the isoelectric point of the proteins [10,11].…”

Section: Introductionmentioning

confidence: 99%

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Microcalorimetric study of the adsorption of PEGylated lysozyme on a strong cation exchange resin

Blaschke

Varon

Werner

et al. 2011

Journal of Chromatography A

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Section: Isothermal Titration Calorimetrymentioning

confidence: 99%

Section: Evaluation Of Itc Experimentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microcalorimetric study of the adsorption of PEGylated lysozyme on a strong cation exchange resin

Blaschke

Varon

Werner

et al. 2011

Journal of Chromatography A

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“…Combining this with the Langmuir isotherm, the equilibrium constant can be calculated as q approaches zero, 15,32…”

Section: Calorimetric Measurements Of Lysozyme Adsorption On Ha-smentioning

confidence: 99%

“…ITC has been successfully used to acquire thermodynamic data in protein adsorption on various ion exchange and hydrophobic interaction adsorbents. [13][14][15] For this purpose, we synthesized a histamine-Sepharose (HA-S) gel with higher ligand density than the histamine-ligand chromatography media reported previously. [16][17][18] Lysozyme was chosen for adsorption onto HA-S because it has been used extensively as a model protein for fundamental researches on chromatographic techniques.…”

Section: Introductionmentioning

confidence: 99%

Studies of lysozyme binding to histamine as a ligand for hydrophobic charge induction chromatography

Shi

Shen

Sun

2009

Biotechnology Progress

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Histamine was immobilized on Sepharose CL-6B (Sepharose) for use as a ligand of hydrophobic charge induction chromatography (HCIC) of proteins. Lysozyme adsorption onto Histamine-Sepharose (HA-S) was studied by adsorption equilibrium and calorimetry to uncover the thermodynamic mechanism of the protein binding. In both the experiments, the influence of salt (ammonium sulfate and sodium sulfate) was examined. Adsorption isotherms showed that HA-S exhibited a high salt tolerance in lysozyme adsorption. This property was well explained by the combined contributions of hydrophobic interaction and aromatic stacking. The isotherms were well fitted to the Langmuir equation, and the equilibrium parameters for lysozyme adsorption were obtained. In addition, thermodynamic parameters (DeltaH(ads), DeltaS(ads), and DeltaG(ads)) for the adsorption were obtained by isothermal titration calorimetry by titrating lysozyme solutions into the adsorbent suspension. Furthermore, free histamine was titrated into lysozyme solution in the same salt-buffers. Compared with the binding of lysozyme to free histamine, lysozyme adsorption onto HA-S was characterized by a less favorable DeltaG(ads) and an unfavorable DeltaS(ads) because histamine was covalently attached to Sepharose via a three-carbon-chain spacer. Consequently, the immobilized histamine could only associate with the residues on the protein surface rather than those in the hydrophobic pocket, causing a less favorable orientation between histamine and lysozyme. Further comparison of thermodynamic parameters indicated that the unfavorable DeltaS(ads) was offset by a favorable DeltaH(ads), thus exhibiting typical enthalpy-entropy compensation. Moreover, thermodynamic analyses indicated the importance of the dehydration of lysozyme molecule and HA-S during the adsorption and a substantial conformational change of the protein during adsorption. The results have provided clear insights into the adsorption mechanisms of lysozyme onto the new HCIC material.

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Recent trends and some applications of isothermal titration calorimetry in biotechnology

Roselin

Lin

et al. 2010

Biotechnology Journal

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Isothermal titration calorimeters (ITCs) are thermodynamic instruments used for the determination of enthalpy changes in any physical/chemical reaction. This can be applied in various fields of biotechnology. This review explains ITC applications, especially in bioseparation, drug development and cell metabolism. In liquid chromatography, the separation/purification of specific proteins or polypeptides in a mixture is usually achieved by varying the adsorption affinities of the different proteins/polypeptides for the adsorbent under different mobile-phase conditions and temperatures. Using ITC analysis, the binding mechanism of proteins with adsorbent solid material is derived by elucidating enthalpy and entropy changes, which offer valuable guidelines for designing experimental conditions in chromatographic separation. The binding affinity of a drug with its target is studied by deriving binding enthalpy and binding entropy. To improve the binding affinity, suitable lead compounds for a drug can be identified and their affinity tested by ITC. Recently ITC has also been used in studying cell metabolism. The heat produced by animal cells in culture can be used as a primary indicator of the kinetics of cell metabolism, which provides key information for drug bioactivity and operation parameters for process cell culture.

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Microcalorimetric study of adsorption of human monoclonal antibodies on cation exchange chromatographic materials

Cited by 30 publications

References 31 publications

Microcalorimetric study of the adsorption of PEGylated lysozyme on a strong cation exchange resin

Microcalorimetric study of the adsorption of PEGylated lysozyme on a strong cation exchange resin

Studies of lysozyme binding to histamine as a ligand for hydrophobic charge induction chromatography

Recent trends and some applications of isothermal titration calorimetry in biotechnology

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