| 1Intact & 5% impurity |
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Applications
Interferon Alpha intact 5% impurity on Jupiter 3u C18 and Jupiter 5u C4
18073
Separation Mode: Reversed Phase
Reversed Phase
C18
HPLC
Pharmaceutical/Biopharmaceutical
Life Science
Clinical Research
Therapeutic / Clinical
Analytes
Details
LC Conditions (App ID: 18073)
Column: |
Brand Name: Jupiter |
Part No: |
Phase Name: C18 |
Sample Note: Application Focus: To demonstrate Jupiter 300 utility for separating folded from unfolded biogeneric proteins
Unlike other separation techniques, reversed phase can often visualize differences between intact and unfolded/ mis-folded protein states. Especially with E.Coli produced recombinant proteins, refolding analysis is often required as part of both manufacturing process analysis technology as well as quality control testing of a finished product. In this example Interferon alpha was exposed to chemical unfolding conditions at higher temperature in the presence of DTT (3 mM dithiothreitol (DTT) at 55°C for 30 min) and the chromatographic behavior of the unfolded protein was compared to the intact protein by analysis on the Jupiter 300 3u C18 and Jupiter 300 5u C4 columns.
In App ID# 18073 5% of unfolded interferon was added to the intact protein. As one can see from the overlaid chromatograms, both columns could easily detect unfolded impurities lower than 5% with good, rapid resolution from the intact protein in fewer than 15 minutes. This application demonstrates the utility of Jupiter 300 media in doing refold analysis. |
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Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide
Phenomenex is a technology leader committed to developing novel analytical chemistry solutions that solve the separation and purification challenges of researchers worldwide