Chromatography Media

doi:10.1002/9783527630158.ch3

Cited by 10 publications

(2 citation statements)

References 62 publications

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“…An exact description of changes that result from a pressure rise are for such separations difficult, due to a constant change in the mobile phase composition. As ion exchange chromatography became one of the most used techniques for protein separation, and gradient separations became necessary to compensate for the steep adsorption isotherms, a model for linear gradient elution (LGE-IEX) prediction was established. − We decided to test the application of the LGE model for the evaluation of the partial molar volume changes as well as changes in the number of binding sites of oligonucleotides and proteins at pressures up to 550 bar while keeping constant temperature and flow rate of the mobile phase. The oligonucleotides Δ V and the number of binding sites were compared to values obtained from description of isocratic separations by stoichiometric displacement model (SDM).…”

Section: Introductionmentioning

confidence: 99%

Effect of Pressure Increase on Macromolecules’ Adsorption in Ion Exchange Chromatography

et al. 2020

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In this study a new method for evaluating the pressure effect on separations of oligonucleotides and proteins on an anion exchange column was developed. The pressure rise of up to 500 bar was attained by coupling restriction capillaries to the column outlet to minimize differences in pressure over the column. Using pH transient measurements it was demonstrated that no shift in ion exchange equilibria occurs due to a pressure increase. Results from isocratic and gradient separations of oligonucleotides (model compounds) were evaluated by stoichiometric displacement and linear gradient elution model, respectively. Both elution modes demonstrated that for smaller oligonucleotides the number of binding sites remained unchanged with pressure rise while an increase for large oligonucleotides was observed, indicating their alignment over the stationary phase. From the obtained model parameters and their pressure dependencies, a thermodynamic description was made and compared between the elution modes. A complementary pattern of a linear increase of partial molar volume change with a pressure rise was established. Furthermore, estimation of the pressure effect was performed for bovine serum albumin and thyroglobulin that required gradient separations. Again, a raise in binding site number was found with pressure increase. The partial molar volume changes of BSA and Tg at the maximal investigated pressure and minimal salt concentration were −31.6 and −34.4 cm3/mol, respectively, indicating a higher rigidity of Tg. The proposed approach provides an insight into the molecule deformation over a surface at high pressures under nondenaturing conditions. The information enables a more comprehensive UHPLC method development.

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Section: Introductionmentioning

confidence: 99%

Effect of Pressure Increase on Macromolecules’ Adsorption in Ion Exchange Chromatography

et al. 2020

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“…Those dependencies can be determined by measurements of isotherms for solutes at various modifier concentrations in the mobile phase, and then, the obtained coefficients can be related with the modifier content [50 -52]. These dependencies can also be estimated by acquiring the band profiles of solutes eluted at different gradient steepness and relating their retention with the gradient shape using the inverse method [53,54] or equilibrium theory [55]. Such a method provides good results if the competition between the modifier and solutes for the adsorption sites can be neglected.…”

Section: Short-cut Methods To Determine the Influence Of The Modifier mentioning

confidence: 99%

Thermodynamics of Multicomponent Liquid Chromatography

Antos

Pia̧tkowski

2016

Chemie Ingenieur Technik

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Thermodynamic models of multicomponent adsorption in liquid chromatography are summarized and reviewed. A general description of adsorption equilibrium is presented, which is followed by simplified approaches that can be adopted for practical applications. Synergistic and competitive adsorption effects accompanying chromatographic elution are illustrated and methods of their predictions are indicated. The influence of a mobile phase modifier on the retention behavior of chromatographed species is discussed. A manner of mathematical predictions of competitive adsorption of modifiers and solutes is presented. Also in this case the analysis starts with a thermodynamically grounded general description and ends with simplified models suitable for practical purposes of design and optimization of chromatographic separations.

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Affinity Chromatography: An Enabling Technology for Large‐Scale Bioprocessing

Lacki¹,

Riske²

2019

Biotechnology Journal

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Affinity chromatography (AC) has been used in large‐scale bioprocessing for almost 40 years and is considered the preferred method for primary capture in downstream processing of various types of biopharmaceuticals. The objective of this mini‐review is to provide an overview of a) the history of bioprocess AC, b) the current state of platform processes based on affinity capture steps, c) the maturing field of custom developed bioprocess affinity resins, d) the advantages of affinity capture‐based downstream processing in comparison to other forms of chromatography, and e) the future direction for bioprocess scale AC. The use of AC can result in economic advantages by enabling the standardization of process development and the manufacturing processes and the use of continuous operations in flexible multiproduct production suites. These concepts are discussed from a growing field of custom affinity bioprocess resin perspective. The custom affinity resins not only address the need for a capture resin for non‐platformable processes, but also can be employed in polishing applications, where they are used to define and control drug substance composition by separating specific product variants from the desired product form.

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Chromatography Media

Cited by 10 publications

References 62 publications

Effect of Pressure Increase on Macromolecules’ Adsorption in Ion Exchange Chromatography

Effect of Pressure Increase on Macromolecules’ Adsorption in Ion Exchange Chromatography

Thermodynamics of Multicomponent Liquid Chromatography

Affinity Chromatography: An Enabling Technology for Large‐Scale Bioprocessing

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