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Related Concept Videos

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
When the...
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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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Diffusion on Chromatography Columns01:07

Diffusion on Chromatography Columns

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In column chromatography, when an analyte is introduced as a narrow band at the top of the column, the solutes begin to separate and broaden, developing a Gaussian profile. This broadening occurs due to various factors, such as longitudinal diffusion.
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Silica Gel Column Chromatography: Overview01:10

Silica Gel Column Chromatography: Overview

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Silica gel column chromatography is a technique for separating compounds using a column packed with silica gel as the stationary phase. This method relies on differences in the polarity of compounds. Based on their polarities, compounds move between the stationary phase (silica gel) and the mobile phase (the solvent), forming discrete bands in the column.
Polar components tend to bind strongly to the silica gel, causing them to move slowly through the column. In contrast, nonpolar compounds...
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Gas Chromatography: Types of Columns and Stationary Phases01:17

Gas Chromatography: Types of Columns and Stationary Phases

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Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...
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Muscles of the Vertebral Column01:27

Muscles of the Vertebral Column

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The back muscles that lie deep into the thoracolumbar fascia are called intrinsic or true back muscles. These muscles are divided into four layers: superficial, intermediate, deep, and deepest layers.
Superficial Layer:
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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
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A Direct Approach for Process Development Using Single Column Experiments Results in Predictable Streamlined

Aditya Utturkar1, Keith Gillette1, Chia-Yun Sun1

  • 1Pall Corporation, 20 Walkup Drive, Westborough, MA, 01581, USA.

Biotechnology Journal
|October 6, 2018
PubMed
Summary

This study developed a streamlined downstream process for monoclonal antibody (mAb) purification using continuous chromatography. The novel approach rapidly optimized parameters, enabling efficient mAb recovery and high purity in just seven weeks.

Keywords:
bioprocess developmentchromatographydownstream Processingprotein purification

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Area of Science:

  • Biopharmaceutical Manufacturing
  • Chemical Engineering
  • Process Development

Background:

  • Continuous monoclonal antibody (mAb) bioprocessing requires efficient downstream process development.
  • Traditional methods can be time-consuming and resource-intensive.

Purpose of the Study:

  • To develop a streamlined, three-step continuous chromatography process for mAb purification.
  • To optimize process parameters using a design of experiment (DoE) approach.
  • To accurately transfer batch process knowledge to a continuous mode of operation.

Main Methods:

  • Utilized a design of experiment (DoE) approach in single-column (batch) mode to simulate multi-column (continuous) purification.
  • Characterized Protein A capture, anion exchange, and mixed-mode cation exchange chromatography steps.
  • Employed empirical modeling to define multi-column chromatography capacity and facilitate batch-to-continuous transfer.

Main Results:

  • Developed two robust and predictable continuous bioprocesses within seven weeks.
  • Achieved a total product recovery of at least 74%.
  • Ensured host cell protein (HCP) content below 5 ppm and aggregate content below 1%.

Conclusions:

  • The implemented process development strategy effectively mimics continuous operation.
  • This approach offers flexibility for developing multiple continuous processes with targeted mAb recovery and purity.
  • The batch-to-continuous transfer methodology is efficient and reliable for biopharmaceutical manufacturing.