Related Experiment Videos
Calmodulin as a versatile tag for antibody fragments
1Cambridge Centre for Protein Engineering, MRC Centre, UK.
Bio/Technology (Nature Publishing Company)
|April 1, 1995
Summary
Researchers developed a versatile calmodulin tag for antibody fragments, enabling efficient bacterial expression and purification. This tag facilitates protein detection and heterodimerization, advancing antibody engineering applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Engineering
Background:
- Calmodulin is an acidic protein with calcium-dependent high-affinity binding to ligands.
- These properties make calmodulin a suitable candidate for protein tagging applications.
Purpose of the Study:
- To develop calmodulin as a versatile tag for antibody fragments.
- To explore its utility in protein purification, detection, and heterodimerization.
Main Methods:
- Constructing calmodulin-single chain Fv fragment (scFv) fusions.
- Expressing and purifying fusions from bacterial sources.
- Utilizing calmodulin's ligand-binding properties for purification and detection.
Main Results:
- High yield secretion of calmodulin-scFv fusions from bacteria (5-15 mg/l).
- Single-step purification via anion-exchange or calmodulin-specific ligand affinity chromatography.
- Successful detection using fluorescently labeled peptide ligands in various assays.
- Demonstrated protein heterodimerization capability.
Conclusions:
- Calmodulin is an effective and versatile tag for antibody fragments.
- Calmodulin tagging enables efficient expression, purification, and detection of antibody fragments.
- This approach facilitates protein heterodimerization for biotechnological applications.