Exploration and Modulation of Antibody Fragment Biophysical Properties by Replacing the Framework Region Sequences
Abstract
:1. Introduction
2. Results
2.1. Biophysical Properties of Wild-Type scFv
2.2. Methodology for Replacing the Framework (FR) Sequences
2.3. Functional Analysis
3. Discussion
4. Materials and Methods
4.1. Protein Expression and Purification
4.2. Biochemical Characterization and scFv Integrity Analysis
4.3. Determination of Thermal and Chemical Stabilities
4.4. Affinity Analysis by Surface Plasmon Resonance
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations/Acronyms
AA | Amino acid |
CDR | complementary-determining regions |
FR | Framework |
Gly | glycine |
GdnHCl | guanidinium chloride |
PBS | phosphate-buffered saline |
scFv | single-chain antibody variable fragment |
SDS-PAGE | sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
SEC | size exclusion chromatography |
SPR | surface plasmon resonance |
Tm | midpoint temperature |
VH | heavy-chain variable |
VL | light-chain variable |
WT | Wild-type |
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Cnudde, T.; Lakhrif, Z.; Bourgoin, J.; Boursin, F.; Horiot, C.; Henriquet, C.; di Tommaso, A.; Juste, M.O.; Jiacomini, I.G.; Dimier-Poisson, I.; et al. Exploration and Modulation of Antibody Fragment Biophysical Properties by Replacing the Framework Region Sequences. Antibodies 2020, 9, 9. https://0-doi-org.brum.beds.ac.uk/10.3390/antib9020009
Cnudde T, Lakhrif Z, Bourgoin J, Boursin F, Horiot C, Henriquet C, di Tommaso A, Juste MO, Jiacomini IG, Dimier-Poisson I, et al. Exploration and Modulation of Antibody Fragment Biophysical Properties by Replacing the Framework Region Sequences. Antibodies. 2020; 9(2):9. https://0-doi-org.brum.beds.ac.uk/10.3390/antib9020009
Chicago/Turabian StyleCnudde, Thomas, Zineb Lakhrif, Justine Bourgoin, Fanny Boursin, Catherine Horiot, Corinne Henriquet, Anne di Tommaso, Matthieu Olivier Juste, Isabella Gizzi Jiacomini, Isabelle Dimier-Poisson, and et al. 2020. "Exploration and Modulation of Antibody Fragment Biophysical Properties by Replacing the Framework Region Sequences" Antibodies 9, no. 2: 9. https://0-doi-org.brum.beds.ac.uk/10.3390/antib9020009