Seeking an explanation of which protein purification method is more efficient and maintains higher protein activity? Thank you!
There are many methods for protein purification, and choosing the appropriate method depends on the characteristics, source, and purpose of the target protein. An efficient protein purification method requires obtaining high purity while maintaining protein activity. Here are some commonly used purification methods that can achieve efficient purification and maintain protein activity to varying degrees:
1. Affinity chromatography:
This is a highly efficient purification method that usually maintains protein activity. By utilizing the highly specific interaction between the target protein and a specific ligand, highly purified protein can be obtained in a single step. Common affinity tags include His-tag, GST-tag, and MBP-tag.
2. Ion exchange chromatography:
Ion exchange chromatography separates proteins based on their net charge at a given pH condition. Selecting appropriate buffer conditions can achieve efficient purification while maintaining protein activity.
3. Size exclusion chromatography (SEC):
Size exclusion chromatography separates proteins based on molecular size and is generally considered a gentle purification method because it does not subject proteins to extreme chemical environments, helping to maintain protein activity. However, SEC is typically used as part of a combination purification strategy, rather than as a sole purification method.
4. Hydrophobic interaction chromatography (HIC):
Hydrophobic interaction chromatography separates proteins based on their hydrophobicity. Appropriate buffer conditions can maintain protein activity and achieve effective purification. However, overly hydrophobic conditions may cause protein inactivation or aggregation.
The choice of an efficient purification method that maintains protein activity depends on the characteristics and requirements of the target protein. Affinity chromatography is often the preferred method as it can achieve high purification and maintain activity in a single step. Other methods can be combined based on the properties and purification needs of the protein.
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Protein-protein interaction mass spectrometry analysis
Protein purity analysis (size exclusion/reverse-phase chromatography)
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