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Related Concept Videos

Ion Exchange01:17

Ion Exchange

Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or basic...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Analyte Adsorption and Distribution01:09

Analyte Adsorption and Distribution

In certain chromatographic separations, solutes transfer between the mobile phase and the stationary phase via sorption, which typically refers to the process of adsorption. For many chromatographic systems, the sorption process often depends on the polarity of the compounds—an expression of the overall dipole moment within the molecule. During the separation process, there is competition between the solute and solvent for adsorption to the stationary phase. Highly polar compounds and solvents...
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:

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Corrigendum to "In-line spectroscopic measurement of pH profiles using methyl orange: Application to pH transients in protein A chromatography" [Journal of Chromatography A 1782 (2026) 467106].

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Related Experiment Video

Updated: May 29, 2026

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
10:41

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Published on: April 5, 2019

Mobile phase modifier effects in multimodal cation exchange chromatography.

Melissa A Holstein1, Siddharth Parimal, Scott A McCallum

  • 1Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180, USA.

Biotechnology and Bioengineering
|September 8, 2011
PubMed
Summary
This summary is machine-generated.

Multimodal chromatography shows distinct protein binding compared to ion exchange. Mobile phase modifiers like arginine significantly alter protein adsorption and selectivity in these systems.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Separation Science

Background:

  • Multimodal chromatography offers unique protein binding capabilities.
  • Understanding mobile phase modifier effects is crucial for optimizing separations.

Purpose of the Study:

  • To investigate protein adsorption in multimodal chromatography.
  • To evaluate the impact of mobile phase modifiers on protein binding and selectivity.
  • To elucidate the structural basis of protein-multimodal ligand interactions.

Main Methods:

  • Utilized multimodal and ion exchange chromatography with a diverse protein library.
  • Employed Nuclear Magnetic Resonance (NMR) spectroscopy, including Saturation Transfer Difference (STD) NMR.
  • Analyzed the effects of various mobile phase modifiers (arginine, guanidine, sodium caprylate, polyols).

Main Results:

  • Multimodal resins exhibit different protein binding behavior and selectivity compared to ion exchange resins.
  • Arginine and guanidine dramatically alter protein adsorption and selectivity.
  • Sodium caprylate can significantly reduce retention for specific proteins, with competitive binding confirmed via STD NMR.
  • Polyols modulate retention based on protein binding strength, balancing electrostatic and hydrophobic interactions.

Conclusions:

  • Protein adsorption in multimodal chromatography is distinct and influenced by mobile phase composition.
  • Mobile phase modifiers offer powerful tools to tune selectivity in protein separations.
  • This study enhances understanding for developing advanced protein separation technologies.