Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

2.7K
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...
2.7K
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

1.7K
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...
1.7K
Analyte Adsorption and Distribution01:09

Analyte Adsorption and Distribution

2.4K
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...
2.4K
Centrifugation01:05

Centrifugation

6.4K
Centrifugation is a separation technique based on differences in density or size. It is commonly used to separate solids from aqueous interferents. During centrifugation, the sample is placed in centrifugation tubes and spun at high angular velocity, which allows centrifugal force to act differentially on the different densities or masses of the components. After spinning, the supernatant liquid is decanted. Depending on the specific application, either the pellet or the supernatant is retained...
6.4K
Types Of Column Chromatography01:29

Types Of Column Chromatography

13.1K
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.
Gel Filtration Chromatography
When the...
13.1K
Enzyme Kinetics01:19

Enzyme Kinetics

103.4K
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
103.4K

You might also read

Related Articles

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

Sort by
Same author

Virtual special issue for HPLC 2024.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences·2026
Same author

Studies and analysis of drug-target interactions by affinity chromatography and related techniques: A review.

Journal of pharmaceutical analysis·2026
Same author

Screening of binding by antidiabetic drugs to normal vs AGE-modified human serum albumin through covalent immobilization and microscale affinity chromatography.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences·2026
Same author

Microscale Affinity Chromatography for Biointeraction Analysis: Strategies, Principles and Applications.

Journal of separation science·2026
Same author

Chromatographic-Based Binding and Thermodynamic Studies of Antibiotic Micropollutants with Humic Acid Using Affinity Microcolumns.

Journal of separation science·2026
Same author

Refreshing the Aims and Scope of the Journal of Chromatography B.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences·2025

Related Experiment Video

Updated: Jan 4, 2026

The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis
19:16

The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis

Published on: March 17, 2010

21.0K

Kinetic Analysis by Affinity Chromatography.

Sazia Iftekhar1, Susan T Ovbude1, David S Hage1

  • 1Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE, United States.

Frontiers in Chemistry
|November 5, 2019
PubMed
Summary
This summary is machine-generated.

This review explores affinity chromatography techniques for studying biological reaction kinetics. It details methods like band-broadening and peak analysis to determine rate constants in living systems.

Keywords:
affinity chromatographybiological interactionskineticspeak decay methodpeak profilingplate height methodsplit-peak methodultrafast affinity extraction

More Related Videos

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

3.5K
Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface
13:22

Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

Published on: November 2, 2011

15.3K

Related Experiment Videos

Last Updated: Jan 4, 2026

The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis
19:16

The Importance of Correct Protein Concentration for Kinetics and Affinity Determination in Structure-function Analysis

Published on: March 17, 2010

21.0K
Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
07:19

Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein

Published on: April 26, 2024

3.5K
Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface
13:22

Adhesion Frequency Assay for In Situ Kinetics Analysis of Cross-Junctional Molecular Interactions at the Cell-Cell Interface

Published on: November 2, 2011

15.3K

Area of Science:

  • Biochemistry and Chemical Kinetics
  • Biophysical Chemistry
  • Analytical Chemistry

Background:

  • Understanding biological reaction rates is crucial for elucidating chemical processes in living systems.
  • Affinity chromatography is a powerful separation technique with applications in kinetic studies.

Purpose of the Study:

  • To review various affinity chromatography techniques for examining biological reaction kinetics.
  • To discuss the principles, theory, applications, advantages, and limitations of these methods for determining rate constants.

Main Methods:

  • Traditional and high-performance affinity chromatography.
  • Techniques discussed include band-broadening measurements, peak fitting, split-peak analysis, peak decay studies, and ultrafast affinity extraction.

Main Results:

  • Detailed discussion of the principles and theory behind each method for rate constant determination.
  • Consideration of the practical applications, advantages, and limitations of each technique.

Conclusions:

  • Affinity chromatography offers a versatile toolkit for investigating the kinetics of biological interactions.
  • These methods provide valuable insights into the rates of biochemical processes, aiding in a deeper understanding of living systems.