Solution-Based Determination of Dissociation Constants for the Binding of Aβ42 to Antibodies

ChemistryOpen
Tao ZhangDieter Willbold

Abstract

Amyloid β-peptides (Aβ) play a major role in the pathogenesis of Alzheimer's disease. Therefore, numerous monoclonal antibodies against Aβ have been developed for basic and clinical research. The present study applied fluorescence based analytical ultracentrifugation and microscale thermophoresis to characterize the interaction between Aβ42 monomers and three popular, commercially available antibodies, namely 6E10, 4G8 and 12F4. Both methods allowed us to analyze the interactions at low nanomolar concentrations of analytes close to their dissociation constants (KD) as required for the study of high affinity interactions. Furthermore, the low concentrations minimized the unwanted self-aggregation of Aβ. Our study demonstrates that all three antibodies bind to Aβ42 monomers with comparable affinities in the low nanomolar range. KD values for Aβ42 binding to 6E10 and 4G8 are in good agreement with formerly reported values from SPR studies, while the KD for 12F4 binding to Aβ42 monomer is reported for the first time.

References

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Citations

Nov 3, 2020·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Sowmya IndrakumarGünther H J Peters

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Methods Mentioned

BETA
surface
isothermal titration calorimetry
ELISA
chip
microscale thermophoresis
glycosylation
chips

Software Mentioned

Sedphat
Sednterp
OriginPro
GUSSI
Sedfit

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