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Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag
Published on: January 16, 2012
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An Orthogonal Fusion Tag for Efficient Protein Purification
Johan Nilvebrant1, Mikael Åstrand1, Sophia Hober2
1Division of Protein Technology, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH/AlbaNova University Center, Stockholm, Sweden.
Methods in Molecular Biology (Clifton, N.J.)
|October 31, 2020
Summary
We developed ABDz1, a dual affinity tag for efficient protein purification. This method uses two specific binding steps for high-purity protein isolation with standard lab equipment.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Chemistry
Background:
- Protein purification is crucial for biochemical and molecular biology research.
- Existing methods may lack specificity or efficiency.
- Novel affinity tags are needed to improve purification processes.
Purpose of the Study:
- To present an efficient method for stringent protein purification.
- To introduce a novel dual affinity tag, ABDz1.
- To demonstrate the utility of ABDz1 in protein purification.
Main Methods:
- Development of a dual affinity tag (ABDz1) based on an albumin-binding domain.
- Utilizing orthogonal affinity purification based on native binding to human serum albumin and engineered binding to Staphylococcal Protein A.
- Fusion of the ABDz1 tag to either terminus of the protein of interest.
Main Results:
- The ABDz1 tag enables a two-step orthogonal affinity purification approach.
- High specificity and efficiency in protein purification were achieved.
- The method is compatible with standard laboratory equipment.
Conclusions:
- The ABDz1 dual affinity tag provides an efficient and stringent method for protein purification.
- This approach simplifies protein purification workflows.
- ABDz1 is a valuable tool for researchers in molecular biology and biochemistry.
