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Related Concept Videos

Protein-Drug Binding: Determination Methods01:22

Protein-Drug Binding: Determination Methods

Determining protein-drug binding can be achieved through indirect and direct methods, each providing valuable insights into the interaction between proteins and drugs.
Indirect methods involve isolating the bound drug from its free form in biological samples such as blood, serum, or plasma. These techniques aim to measure the percentage of drugs bound to proteins. Equilibrium dialysis is a commonly used method where the free drug concentration at equilibrium is measured by separating the bound...
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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
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Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples

Published on: May 4, 2012

Microarray-based study of carbohydrate-protein binding.

Zhenxin Wang1, Jingqing Gao

  • 1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, and Graduate School of the Chinese Academy of Sciences, Beijing, China.

Methods in Molecular Biology (Clifton, N.J.)
|November 3, 2009
PubMed
Summary
This summary is machine-generated.

A new high-throughput microarray assay enables monitoring of carbohydrate-protein interactions. This tool uses fluorescent dyes and gold nanoparticles for sensitive detection of lectin binding to carbohydrates and glycoproteins.

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Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Carbohydrate-protein interactions are crucial in biological processes.
  • Developing high-throughput methods for studying these interactions is essential.
  • Existing methods may lack sensitivity or throughput for comprehensive analysis.

Purpose of the Study:

  • To create a novel, high-throughput microarray tool for monitoring carbohydrate-protein interactions.
  • To establish a sensitive and selective assay for detecting lectin binding.
  • To validate the assay using well-defined carbohydrate and glycoprotein recognition systems.

Main Methods:

  • Preparation of carbohydrate or glycoprotein microarrays.
  • Utilizing avidin-biotin chemistry for fluorescent dye and gold nanoparticle attachment.
  • Employing fluorescence and resonance light scattering (RLS) detection.
  • Enhancing RLS signals via silver electroless deposition onto gold nanoparticles.

Main Results:

  • Demonstrated highly selective recognition of carbohydrate-protein interactions.
  • Achieved sensitive detection limits: 25.6 pg/mL for RCA120 (lectin) in solution.
  • Quantified specific binding on microarray spots: 8 µM for Gal-beta (monosaccharide) and 32 ng/mL for Asf (glycoprotein).

Conclusions:

  • The developed microarray assay is a powerful tool for high-throughput monitoring of carbohydrate-protein interactions.
  • The assay demonstrates high sensitivity and selectivity, suitable for various biological studies.
  • This method offers a valuable platform for glycomics and drug discovery research.