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Related Concept Videos

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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Ion-Exchange Chromatography01:09

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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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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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Optimizing Chromatographic Separations01:15

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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.
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High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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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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Principles Of Column Chromatography01:13

Principles Of Column Chromatography

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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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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, R Bhambure2

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

Methods in Molecular Biology (Clifton, N.J.)
|October 31, 2020
PubMed
Summary
This summary is machine-generated.

High-throughput process development (HTPD) offers a systematic approach to optimize chromatographic separation, overcoming challenges in biopharmaceutical manufacturing. This method integrates miniaturization and automation for efficient and reliable process development.

Keywords:
Design of experiments (DOE)Design spaceHigh-throughput process development (HTPD)Ion-exchange chromatography (IEX)MiniaturizationPreDictor™ plates

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Area of Science:

  • Biopharmaceutical Process Development
  • Chromatographic Separation Science
  • High-Throughput Experimentation

Background:

  • Chromatographic separation is crucial for purifying biopharmaceuticals, removing various impurities.
  • Traditional chromatographic process development faces challenges like complex raw materials, low concentrations, and poor mechanistic understanding, often resulting in suboptimal processes.
  • Time and resource constraints in the biopharma industry necessitate more efficient development strategies.

Purpose of the Study:

  • To describe a protocol for high-throughput process development (HTPD) of chromatography steps.
  • To present a systematic approach for time- and resource-efficient chromatographic process development.
  • To demonstrate the utility of HTPD in generating data representative of traditional lab-scale experiments.

Main Methods:

  • Utilized a commercially available HTPD platform (PreDictor™ plates) in a 96-well format.
  • Integrated miniaturization, automation, and parallelization for systematic process development.
  • Employed statistical tools for data analysis and incorporated mechanistic process knowledge.

Main Results:

  • The HTPD protocol successfully generated data comparable to traditional lab-scale chromatography.
  • A case study on ion exchange chromatography of Granulocyte Colony Stimulating Factor (GCSF) showed significant data agreement (regression coefficient 0.93).
  • Addressed and provided solutions for challenges encountered during HTPD experiments.

Conclusions:

  • HTPD platforms offer a valuable, systematic, and efficient approach for chromatographic process development in biopharmaceuticals.
  • The described protocol and statistical analysis provide a robust method for optimizing chromatographic steps.
  • This approach is highly relevant for the biopharma industry facing time and resource limitations.