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Related Concept Videos

Optimizing Chromatographic Separations01:15

Optimizing Chromatographic Separations

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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...
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High-Performance Liquid Chromatography: Elution Process01:05

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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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Chromatographic Methods: Terminology01:18

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Chromatography is an analytical technique widely used in fields such as chemistry, biology, environmental science, and pharmaceuticals to separate the components of a mixture and identify substances between them. The process of chromatography is based on the interactions between two distinct phases: the stationary phase and the mobile phase. The stationary phase is fixed in place by a supporting material, while the mobile phase moves over it, carrying the solutes. As the mobile phase travels,...
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Principles Of Column Chromatography01:13

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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...
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Chromatographic Resolution01:15

Chromatographic Resolution

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In chromatography, a solute moves through a chromatographic column and tends to spread, forming a Gaussian-shaped band. The longer the solute spends in the column, the broader the band becomes. The broadening can lead to overlaps within the column, affecting separation effectiveness.
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In chromatography,...
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High-Performance Liquid Chromatography: Instrumentation00:57

High-Performance Liquid Chromatography: Instrumentation

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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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Updated: Aug 29, 2025

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Small, smaller, smallest: Miniaturization of chromatographic process development.

Tiago Castanheira Silva1, Michel Eppink2, Marcel Ottens1

  • 1Department of Biotechnology, Delft University of Technology, van der Maasweg 9, Delft, 2629 HZ, The Netherlands.

Journal of Chromatography. A
|September 9, 2022
PubMed
Summary

A novel microfluidic chip enables efficient adsorption isotherm determination for biopharmaceutical purification, significantly reducing liquid and resin volumes. This cost-effective miniaturization is ideal for expensive therapeutics like monoclonal antibodies.

Keywords:
Batch adsorption protein isothermsHigh-Throughput ScreeningLiquid-Handling StationsMicrofluidic chromatographyMiniaturization

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Area of Science:

  • Biotechnology
  • Chemical Engineering
  • Biopharmaceutical Manufacturing

Background:

  • Therapeutic proteins, including monoclonal antibodies (mAb), are crucial for treating diseases like cancer and autoimmune disorders.
  • Post-cell culture biopharmaceutical production involves significant impurities, necessitating purification steps like chromatography.
  • Current chromatographic process development is costly, time-consuming, and resource-intensive, driving the need for more efficient methods.

Purpose of the Study:

  • To present a microfluidic chip approach for rapid and accurate adsorption isotherm determination in biopharmaceutical purification.
  • To compare the performance and cost-effectiveness of the microfluidic chip against robotic liquid handling and manual methods.
  • To evaluate the impact of miniaturization and automation on chromatographic process development for different product values.

Main Methods:

  • Development and implementation of a sophisticated microfluidic chip for adsorption isotherm studies.
  • Utilization of image analysis software for precise determination of chromatographic resin volume.
  • Comparative analysis of the microfluidic chip against a robotic Liquid-handling Station (LHS) and Eppendorf tubes.

Main Results:

  • The microfluidic chip achieved a 15-fold reduction in liquid volume and a 200/100-fold reduction in resin consumption.
  • Adsorption isotherm performance of the microfluidic chip was comparable to miniaturized robotic and manual techniques.
  • Microfluidic chip-based development was most cost-effective for high-value products (e.g., monoclonal antibodies), while automation (LHS) was preferred for lower-value products (e.g., lysozyme).

Conclusions:

  • Microfluidic chip technology offers a highly efficient and cost-effective solution for chromatographic process development in biopharmaceutical manufacturing.
  • Miniaturization via microfluidics significantly reduces resource consumption and development time, particularly for expensive therapeutic proteins.
  • The choice between microfluidic miniaturization and robotic automation depends on the economic value of the target biopharmaceutical product.