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Related Concept Videos

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Affinity Chromatography

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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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High-Performance Liquid Chromatography: Introduction01:11

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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.
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High-Performance Liquid Chromatography: Instrumentation00:57

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High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.
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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.
Gel Filtration Chromatography
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High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

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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...
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High-Performance Liquid Chromatography: Types of Detectors01:15

High-Performance Liquid Chromatography: Types of Detectors

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The role of the detectors in High-Performance Liquid Chromatography (HPLC) is to analyze the solutes as they exit from the chromatographic column. The detector recognizes the solute's property and generates corresponding electrical signals, which are converted into a readable graph of the detector's response versus elution time called a chromatogram at the computer. There are several types of HPLC detectors, each with its own advantages and limitations, depending on the analyte...
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A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
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High-performance lectin affinity chromatography.

Yuka Kobayashi1

  • 1J-Oil Mills, Inc., 11, Kagetoricho, Totsuka-ku, Yokohama, Kanagawa, 245-0064, Japan, yuka.kobayashi@j-oil.com.

Methods in Molecular Biology (Clifton, N.J.)
|August 14, 2014
PubMed
Summary
This summary is machine-generated.

Lectin high-performance liquid chromatography (HPLC) enables glycoprotein separation and purification. This method, using Lens culinaris agglutinin (LCA) columns, can isolate specific glyco-isomers, such as the liver cancer biomarker α-fetoprotein.

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Area of Science:

  • Glycomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Glycoproteomics research is expanding, driven by advancements in lectin high-performance liquid chromatography (HPLC).
  • Affinity chromatography is a powerful technique for isolating and purifying trace amounts of biological molecules.

Purpose of the Study:

  • To outline a fundamental protocol for separating glycoproteins utilizing commercially available lectin-HPLC columns.
  • To demonstrate the utility of lectin-HPLC in analyzing specific glycoprotein glyco-isomers.

Main Methods:

  • Utilized lectin-HPLC with commercially available columns.
  • Employed Lens culinaris agglutinin (LCA) affinity chromatography.
  • Separated glyco-isomers of α-fetoprotein.

Main Results:

  • Successfully demonstrated the separation of glycoproteins using lectin-HPLC.
  • Showcased the ability to resolve glyco-isomers of α-fetoprotein.
  • Validated the effectiveness of LCA columns for specific glycoprotein analysis.

Conclusions:

  • Lectin-HPLC provides an effective method for glycoprotein separation and purification.
  • The technique allows for the detailed analysis of glycoprotein glyco-isomers.
  • This approach is valuable for biomarker discovery and characterization, exemplified by α-fetoprotein analysis.