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Related Concept Videos

Optimizing Chromatographic Separations01:15

Optimizing Chromatographic Separations

Optimizing chromatographic separations is crucial for obtaining clean separations in a minimum amount of time. Optimization is required for several factors, including kinetic effects related to band broadening, plate height, capacity factor, and separation factor.
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
High-Performance Liquid Chromatography: Elution Process01:05

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...
High-Performance Liquid Chromatography: Introduction01:11

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.
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Principles Of Column Chromatography01:13

Principles Of Column Chromatography

The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
Types Of Column Chromatography01:29

Types Of Column Chromatography

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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Upstream Processing01:27

Upstream Processing

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An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter
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An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter

Published on: May 21, 2021

Model-based high-throughput process development for chromatographic whey proteins separation.

Beckley K Nfor1, Jovana Ripić, Albert van der Padt

  • 1Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.

Biotechnology Journal
|August 14, 2012
PubMed
Summary
This summary is machine-generated.

This study optimized whey protein separation using high-throughput screening (HTS) and chromatography modeling. Capto Q resin proved superior for efficient separation and scale-up, minimizing experimental effort.

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An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Area of Science:

  • Biochemical Engineering
  • Separation Science
  • Chromatography

Background:

  • Whey protein separation is challenging due to similar isoelectric points (pIs).
  • Efficient chromatography resin selection is critical for process development.

Purpose of the Study:

  • To develop and validate an integrated approach for selecting and optimizing chromatography resins for whey protein separation.
  • To compare the performance of anion-exchange chromatography (AEX) and hydrophobic interaction chromatography (HIC) resins.

Main Methods:

  • Employed high-throughput screening (HTS) for initial resin evaluation.
  • Utilized column modeling with batch uptake experiments and isotherm parameter regression.
  • Experimentally validated and optimized chromatographic operations.
  • Scaled up the selected process from laboratory to preparative scale.

Main Results:

  • Capto Q (AEX) demonstrated significantly better performance than Octyl Sepharose 4 FF (HIC) in terms of capacity, loading, throughput, and productivity.
  • The integrated modeling and experimental approach successfully identified optimal operating conditions.
  • Scale-up from 1 mL to 35 L yielded reproducible elution profiles.

Conclusions:

  • The combined HTS and column modeling approach accelerates chromatography process development.
  • Capto Q is an effective resin for industrial-scale whey protein separation.
  • This strategy minimizes experimentation while maximizing the exploration of operating variables.