A Two-Step Approach for the Design and Generation of Nanobodies
Abstract
:1. Introduction
2. Results
2.1. Design of Toxin- and Hapten-Specific VHH Grafts
2.2. Evaluation of Antigen Binding
2.3. Design of a Synthetic VHH Library
2.4. Selection of Fluorescein-Binding VHH from the Synthetic Library
2.5. Characterization of a Selected Fluorescein-Binding VHH
3. Discussion
4. Materials and Methods
4.1. Graft Construction
4.2. Protein Production and Purification
4.3. Synthesis of Flu-BSA
4.4. Analytical Methods
4.5. Synthetic Flu-VHH Library and Phage Display
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
BoNT/A-LC | Botulinum neurotoxin serotype A light chain |
BSA | Bovine serum albumin |
CDR | Complementarity-determining region |
ELISA | Enzyme-linked immunosorbent assay |
EMA | European Medicines Agency |
Fab | Fragment antigen-binding |
FDA | Food and Drug Administration |
Flu | Fluorescein |
HCAb | Heavy-chain-only antibody |
sdAb | Single-domain antibody |
scFv | single-chain variable fragment |
VH | Heavy chain variable domain |
VHH | variable domain of camelid heavy-chain-only antibodies |
VL | Light chain variable domain |
VNAR | Variable domain of the immunoglobulin new antigen receptor (IgNAR) from sharks |
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Wagner, H.J.; Wehrle, S.; Weiss, E.; Cavallari, M.; Weber, W. A Two-Step Approach for the Design and Generation of Nanobodies. Int. J. Mol. Sci. 2018, 19, 3444. https://doi.org/10.3390/ijms19113444
Wagner HJ, Wehrle S, Weiss E, Cavallari M, Weber W. A Two-Step Approach for the Design and Generation of Nanobodies. International Journal of Molecular Sciences. 2018; 19(11):3444. https://doi.org/10.3390/ijms19113444
Chicago/Turabian StyleWagner, Hanna J., Sarah Wehrle, Etienne Weiss, Marco Cavallari, and Wilfried Weber. 2018. "A Two-Step Approach for the Design and Generation of Nanobodies" International Journal of Molecular Sciences 19, no. 11: 3444. https://doi.org/10.3390/ijms19113444