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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
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,...

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Related Experiment Video

Updated: May 11, 2026

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
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Evaluating antibody monomer separation from associated aggregates using mixed-mode chromatography.

Dong Gao1, Li-Li Wang, Dong-Qiang Lin

  • 1Key Laboratory of Synthetic and Natural Functional Molecular Chemistry of Ministry of Education, Institute of Modern Separation Science, Northwest University, Shaanxi Key Laboratory of Modern Separation Science, Xi'an 710068, China. gaod@nwu.edu.cn

Journal of Chromatography. A
|May 4, 2013
PubMed
Summary

Mixed-mode resins effectively remove antibody aggregates, crucial for biopharmaceutical quality. Capto adhere and benzylamine resins significantly reduced aggregates, highlighting the importance of combined molecular interactions.

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Last Updated: May 11, 2026

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Capillary Electrophoresis Separation of Monoclonal Antibody Isoforms Using a Neutral Capillary
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Capillary Electrophoresis Separation of Monoclonal Antibody Isoforms Using a Neutral Capillary

Published on: January 16, 2017

Area of Science:

  • Biopharmaceutical Manufacturing
  • Protein Chemistry
  • Chromatographic Separation

Background:

  • Antibody aggregation impacts product quality and biological activity.
  • Effective separation methods are vital for biopharmaceutical production.
  • Understanding molecular interactions aids in optimizing purification processes.

Purpose of the Study:

  • To evaluate mixed-mode resins for antibody aggregate removal.
  • To compare mixed-mode resins with traditional chromatography resins.
  • To investigate the role of molecular interactions in aggregate separation.

Main Methods:

  • Utilized three mixed-mode resins (Capto adhere, BA, AB) and two traditional resins (Q Sepharose FF, Phenyl Sepharose 6 FF).
  • Assessed separation performance based on aggregate removal efficiency at varying mass loadings.
  • Analyzed the contribution of hydrophobic and electrostatic interactions.

Main Results:

  • Removal efficiency was highly dependent on mass loading.
  • Capto adhere and BA resins significantly reduced aggregate levels from 20.5% to 2.6% and 2.4% at 50mg/ml.
  • Traditional resins showed lower aggregate reduction efficiency.

Conclusions:

  • Mixed-mode resins demonstrate superior performance in antibody aggregate removal.
  • Both hydrophobic and electrostatic interactions are critical for effective aggregate separation.
  • The cooperativity of molecular interactions is key for optimizing mixed-mode resin performance.