Creation of Multimeric Single-Domain Antibodies Using Bacterial Superglues
Paul J Wichgers Schreur1, Sandra van de Water2, Jeroen Kortekaas2,3
1Department of Virology and Molecular Biology, Wageningen Bioveterinary Research, Lelystad, The Netherlands. paul.wichgersschreur@wur.nl.
Methods in Molecular Biology (Clifton, N.J.)
|February 14, 2022
Summary
Creating multimeric antibody molecules is simplified using bacterium-derived superglue components. This method enables straightforward assembly of single-domain antibodies (sdAbs) into advanced antibody formats with enhanced properties.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- Multimerization of single-domain antibodies (sdAbs) enhances avidity, in vivo half-life, and biological activity.
- Traditional methods for antibody multimerization can be complex and time-consuming.
Purpose of the Study:
- To present detailed methods for constructing single-domain antibodies (sdAbs) and scaffolds with genetically fused superglue components.
- To demonstrate the assembly of these components into multimeric antibody complexes.
Main Methods:
- Utilizing two-component superglues derived from bacterial peptides (Tags) and protein domains (Catchers).
- Genetically fusing superglue components to single-domain antibodies (sdAbs) and scaffold proteins.
- Mixing the individual components to facilitate spontaneous isopeptide bond formation and multimer assembly.
Main Results:
- Successful construction of single-domain antibodies (sdAbs) and scaffolds incorporating superglue components.
- Demonstration of efficient assembly into multimeric antibody complexes through simple mixing.
- Establishment of a versatile platform for creating tailored multimeric antibody formats.
Conclusions:
- The superglue system provides a facile and modular approach for constructing multimeric single-domain antibodies (sdAbs).
- This method offers a powerful tool for engineering antibody molecules with improved characteristics for various applications.
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