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Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

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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...
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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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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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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

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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Chromatography: Introduction01:10

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Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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High-throughput process development: I. Process chromatography.

Anurag S Rathore1, Rahul Bhambure

  • 1Department of Chemical Engineering, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India, asrathore@biotechcmz.com.

Methods in Molecular Biology (Clifton, N.J.)
|March 21, 2014
PubMed
Summary
This summary is machine-generated.

High-throughput process development (HTPD) streamlines chromatography by integrating miniaturization and automation. This systematic approach accelerates the development of efficient purification processes for biopharmaceuticals, overcoming traditional trial-and-error limitations.

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Process Development
  • Chromatography

Background:

  • Chromatographic separation is crucial for impurity removal in bioprocessing.
  • Traditional chromatography development faces challenges like complex raw materials and limited understanding, often resulting in suboptimal processes.
  • High-throughput process development (HTPD) offers a systematic solution to these challenges.

Purpose of the Study:

  • To describe a protocol for high-throughput process development (HTPD) of chromatographic steps.
  • To detail the operation of a commercially available HTPD device (PreDictor™ plates).
  • To present a method for statistical analysis of HTPD data and demonstrate its utility.

Main Methods:

  • Utilized a miniaturized, automated, and parallelized HTPD platform (PreDictor™ plates).
  • Integrated mechanistic process knowledge with statistical tools for data analysis.
  • Conducted a case study using ion-exchange chromatography for granulocyte colony-stimulating factor (GCSF).

Main Results:

  • The HTPD protocol generated data highly representative of traditional lab-scale experiments.
  • A significant agreement was observed between HTPD and lab-scale data (regression coefficient 0.93).
  • The study successfully demonstrated the efficiency and accuracy of the HTPD approach.

Conclusions:

  • HTPD provides a time- and resource-efficient method for chromatography process development in the biopharma industry.
  • The described protocol and statistical analysis approach are valuable for optimizing purification processes.
  • This systematic approach overcomes limitations of traditional trial-and-error methods, leading to more robust and efficient bioprocesses.