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Author Spotlight: Optimizing Affinity Chromatography for His-Tagged FEN1 Protein
Published on: April 26, 2024
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Several affinity tags commonly used in chromatographic purification
Xinyu Zhao1, Guoshun Li1, Shufang Liang1
1State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, No. 17, Third Section of Renmin South Road, Chengdu 610041, China.
Journal of Analytical Methods in Chemistry
|February 4, 2014
Summary
This review covers common affinity tags used in proteomics, detailing their pros and cons. It also guides on selecting proteases for effective tag removal to preserve protein function.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Affinity tags are essential tools in biological research for protein purification and detection.
- They aid in isolating proteins of interest and analyzing protein complexes.
- Common tags include hexahistidine, FLAG, Strep II, SBP, CBP, GST, MBP, S-tag, HA, and c-Myc.
Purpose of the Study:
- To review the advantages and disadvantages of frequently used affinity tags.
- To discuss the impact of large affinity tags on protein structure and activity.
- To provide guidance on selecting proteases for tag excision.
Main Methods:
- Literature review of common affinity tags and their properties.
- Analysis of protease characteristics for tag cleavage.
- Comparison of different tag systems and their applications.
Main Results:
- Large tags like GST and MBP can affect fusion protein structure and activity.
- Protease selection is crucial for efficient and specific tag removal.
- Common proteases for tag cleavage include enterokinase, factor Xa, thrombin, TEV, and HRV 3C protease.
Conclusions:
- Choosing the right affinity tag and protease is critical for successful protein studies.
- Understanding tag properties and cleavage mechanisms aids experimental design.
- This review offers a guide for optimizing protein purification and analysis workflows.
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