Rationally Designed Antibodies as Research Tools to Study the Structure–Toxicity Relationship of Amyloid-β Oligomers
Abstract
:1. Introduction
2. Results
2.1. The DesAbs Inhibit Aβ40 Aggregation
2.2. The DesAbs Bind Stabilized Aβ40 Oligomers and Increase Their Hydrophobic Surface Area
2.3. DesAbs Increase the Size of Stabilized Aβ40 Oligomers
2.4. Trodusquemine Binds Stabilized Aβ40 Oligomers and Increases their Hydrophobic Surface Area
2.5. The DesAbs do not Change the Toxicity of Aβ40 Oligomers for Human Neuroblastoma Cells
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Antibody Design, Expression and Purification
4.3. Protein Expression and Chemical Kinetics
4.4. Preparation of the Stabilized Aβ40 Oligomers
4.5. ANS Binding Measurements
4.6. Turbidimetry Measurements
4.7. SLS
4.8. Sedimentataion-Based Dot Blot Assays
4.9. Antibody Binding Dot Blot Assays
4.10. Atomic Force Microscopy
4.11. Neuroblastoma Cell Cultures
4.12. MTT Reduction Assay
4.13. Statistics and Data Availability
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
Aβ | amyloid-β peptide |
Aβ40 | 40-residue form of the amyloid-β peptide |
Aβ42 | 42-residue form of the amyloid-β peptide |
AD | Alzheimer’s disease |
AFM | atomic force microscopy |
ANS | 8-anilinonapthalene-1-sulfonic acid |
DesAb | designed antibody |
HFIP | hexafluoroisopropanol |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
SEM | standard error of the mean |
SLS | static light scattering |
ThT | thioflavin T |
TRO | trodusquemine |
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Limbocker, R.; Mannini, B.; Cataldi, R.; Chhangur, S.; Wright, A.K.; Kreiser, R.P.; Albright, J.A.; Chia, S.; Habchi, J.; Sormanni, P.; et al. Rationally Designed Antibodies as Research Tools to Study the Structure–Toxicity Relationship of Amyloid-β Oligomers. Int. J. Mol. Sci. 2020, 21, 4542. https://doi.org/10.3390/ijms21124542
Limbocker R, Mannini B, Cataldi R, Chhangur S, Wright AK, Kreiser RP, Albright JA, Chia S, Habchi J, Sormanni P, et al. Rationally Designed Antibodies as Research Tools to Study the Structure–Toxicity Relationship of Amyloid-β Oligomers. International Journal of Molecular Sciences. 2020; 21(12):4542. https://doi.org/10.3390/ijms21124542
Chicago/Turabian StyleLimbocker, Ryan, Benedetta Mannini, Rodrigo Cataldi, Shianne Chhangur, Aidan K. Wright, Ryan P. Kreiser, J. Alex Albright, Sean Chia, Johnny Habchi, Pietro Sormanni, and et al. 2020. "Rationally Designed Antibodies as Research Tools to Study the Structure–Toxicity Relationship of Amyloid-β Oligomers" International Journal of Molecular Sciences 21, no. 12: 4542. https://doi.org/10.3390/ijms21124542