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Related Concept Videos

Ion-Exchange Chromatography01:09

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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...
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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...
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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.
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Types Of Column Chromatography01:29

Types Of Column Chromatography

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The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
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Two-dimensional Gel Electrophoresis01:22

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Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
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Gas Chromatography: Types of Columns and Stationary Phases01:17

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Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
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Corrigendum to "Structural determinants of peptide-dependent TAP1-TAP2 transit passage targeted by viral proteins and altered by cancer-associated mutations" [Comput. Struct. Biotechnol. J. 19 (2021) 5072-5091].

Computational and structural biotechnology journal·2022
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CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis
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Different Stationary Phase Selectivities and Morphologies for Intact Protein Separations.

A Astefanei1, I Dapic1, M Camenzuli1

  • 1Centre for Analytical Science in Amsterdam (CASA), Van't Hoff Institute for Molecular Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.

Chromatographia
|May 23, 2017
PubMed
Summary
This summary is machine-generated.

The human proteome is complex, with one gene encoding multiple proteins. This review explores advanced liquid chromatography techniques for improved top-down proteomics, aiding disease research.

Keywords:
Intact proteinsLC–MSLiquid chromatographyTop-down proteomics

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Area of Science:

  • Proteomics
  • Molecular Biology
  • Analytical Chemistry

Background:

  • The central dogma of one gene encoding one protein is an oversimplification; humans have ~20,000 genes encoding numerous protein variants.
  • Post-translational modifications significantly impact protein function, making protein diversity crucial for biological processes.
  • Understanding the human proteome is vital for deciphering disease mechanisms and developing therapeutic strategies.

Purpose of the Study:

  • To review alternative selectivities and stationary phase morphologies in liquid chromatography.
  • To highlight their potential applications in top-down proteomics for analyzing intact proteins.
  • To address the gap in adapting chromatographic advances in proteomics compared to mass spectrometry.

Main Methods:

  • Exploration of advanced liquid chromatography techniques beyond traditional reversed-phase chromatography.
  • Focus on alternative stationary phase morphologies and selectivities.
  • Review of methods applicable to high-resolution and high-sensitivity proteomic analysis.

Main Results:

  • Identified alternative chromatographic approaches with potential for enhanced proteomic analysis.
  • Highlighted the importance of separation techniques for enriching and separating low-abundant proteins.
  • Discussed the need for innovation in chromatography for top-down proteomics.

Conclusions:

  • Advanced liquid chromatography offers promising avenues for improving top-down proteomics.
  • Further adoption of novel chromatographic techniques is essential for comprehensive proteome analysis.
  • Innovations in chromatography can significantly advance our understanding of protein diversity and its role in health and disease.