Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Ion-Exchange Chromatography

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...

High-Performance Liquid Chromatography: Introduction

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:

Size-Exclusion Chromatography

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: Elution Process

High-Performance Liquid Chromatography: Elution Process

In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...

Affinity Chromatography

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...

Extraction: Advanced Methods

Extraction: Advanced Methods

Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is formed in...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Frozen elephant trunk implantation in zone 0 using a left subclavian stented prosthesis.

Multimedia manual of cardiothoracic surgery : MMCTS·2026

Same author

Thirty years of Bose-Einstein condensation.

Nature·2025

Same author

Percentage of left ventricular myocardial blood flow distribution and revascularization completeness in FASTTRACK CABG.

Journal of cardiovascular computed tomography·2025

Same author

High-risk plaques in proximal and distal segments relative to graft anastomoses and non-grafted segments.

Journal of cardiovascular computed tomography·2025

Same author

A supplement about drainology is a compliment to cardiothoracic surgery.

European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery·2025

Same author

Aortic regurgitation in left ventricular assist device patients: Does aortic root dilatation contribute to valve incompetence?

Artificial organs·2024

Search research articles

Related Experiment Video

Updated: Jul 6, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Protein concentration by hydrophilic interaction chromatography combined with solid phase extraction.

Ulrich Schneider¹

¹Siegfried-Biologics GmbH, Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|March 29, 2008

Summary

Hydrophilic interaction chromatography (HILIC) offers a new method for nonselective protein enrichment from dilute samples. This technique enables concentration and buffer exchange directly into a sample buffer for two-dimensional gel electrophoresis (2-D PAGE).

More Related Videos

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

Published on: November 17, 2011

Related Experiment Videos

Last Updated: Jul 6, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

Published on: November 17, 2011

Area of Science:

Biochemistry
Analytical Chemistry
Chromatography

Background:

Hydrophilic interaction chromatography (HILIC) is a normal phase chromatography variant utilizing a high organic modifier mobile phase.
HILIC is established for isolating specific biomolecules like membrane proteins, glycopeptides, and modified protein variants.
Existing HILIC applications require specific elution conditions and may not be suitable for direct sample preparation for subsequent analyses.

Purpose of the Study:

To describe an extended application of HILIC for nonselective protein enrichment from dilute biological sources.
To demonstrate the utility of HILIC for preparing protein samples prior to two-dimensional gel electrophoresis (2-D PAGE).
To introduce a compatible recovery method for simultaneous buffer exchange, concentration, and direct sample loading into 2-D PAGE buffers.

Main Methods:

Utilized hydrophilic interaction chromatography (HILIC) with a poly [2-hydroxyethyl] aspartamide silica stationary phase.
Applied HILIC for the enrichment of proteins from dilute samples, including those from carrier-free electrophoresis (free flow electrophoresis [FFE]).
Developed and demonstrated a compatible recovery technique for direct sample preparation into 2-D PAGE buffer.

Main Results:

Successfully demonstrated nonselective protein enrichment from various dilute sources using HILIC.
Showcased the application of HILIC for protein samples obtained from preparative isoelectric focusing (IEF) separations via FFE.
Validated a one-step compatible recovery process for buffer exchange, concentration, and direct loading into 2-D PAGE sample buffers.

Conclusions:

HILIC can be effectively extended for the nonselective enrichment of proteins from dilute samples.
The developed method simplifies protein sample preparation for downstream analyses like 2-D PAGE.
The compatible recovery technique streamlines the workflow, enabling direct application of enriched proteins into 2-D PAGE.