JoVE Journal

Biology

This content is Open Access.

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

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43.4K Views

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10:21 min

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September 21st, 2011

DOI :

September 21st, 2011

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Patrick J. M. Murphy¹, Orrin J. Stone², Michelle E. Anderson¹

¹College of Nursing, Interdisciplinary Life Sciences Research Laboratory, Seattle University, ²College of Science and Engineering, Interdisciplinary Life Sciences Research Laboratory, Seattle University

Podsumowanie

An automated method for identifying suitable hydrophobic interaction chromatography (HIC) media to be used in the process of protein purification is presented. The method utilizes a medium-pressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multi-wavelength analysis, and split fraction eluate collection.

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Automated Hydrophobic Interaction Chromatography

Protein Purification

Chromatographic Medium Selection

Affinity Chromatography

Ion Exchange Chromatography

Protein Concentration

Ammonium Sulfate

Purification Method

Nonpolar Ligand

Chromatography Column

Salt Concentration

Rozdziały w tym wideo

0:05

Title

1:30

Buffer and Sample Preparation

2:48

Physical Setup and Plumbing of the DuoFlow Chromatography System

4:11

Priming System Lines,Programming Run Method, and Equilibrating HIC Columns

5:43

Sample Loading, Programming the Software, and Running the Column Scouting Protocol

7:36

Representative Chromatogram and Fluorescent Fraction Data

9:23

Conclusion

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