Fluorescent Labeling of Antibody Fragments Using Split GFP
Author Information
Author(s): Ferrara Fortunato, Listwan Pawel, Waldo Geoffrey S., Bradbury Andrew R. M.
Primary Institution: Bioscience Division, Los Alamos National Laboratory, Los Alamos, New Mexico, United States of America
Hypothesis
Can the split green fluorescent protein (GFP) system be used to efficiently label single chain Fvs (scFvs) without affecting their functionality?
Conclusion
The split GFP system allows for effective fluorescent labeling of scFvs without compromising their expression or functionality.
Supporting Evidence
- The GFP11 tag did not affect scFv expression or functionality.
- Fluorescent labeling was demonstrated in various assays including flow cytometry and immunosorbent assays.
- The split GFP system can quantify scFv concentrations in crude samples without purification.
Takeaway
This study shows a new way to make antibodies glow using a special protein that doesn't get in the way of how they work.
Methodology
The study involved genetic fusion of the GFP11 peptide to the anti-chicken-lysozyme scFv, D1.3, and evaluated its expression and functionality in various assays.
Limitations
The study does not address potential long-term stability of the labeled scFvs in various conditions.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website