Figure 1.
Surface preparation with different Aβ(1–42) assembly states for surface plasmon resonance (SPR) analysis.
Size exclusion chromatography (monomers and oligomers) and density gradient centrifugation (fibrils) ensure highly pure samples for immobilization on sensor surfaces and subsequent SPR measurements.
Figure 2.
Interaction of scFv-IC16 with different immobilized Aβ(1–42) assembly states.
SPR sensorgrams were recorded separately with single-cycle kinetics. Experimentally obtained, double-referenced binding data (black traces) were superimposed with the corresponding fit (red traces). Monomer data was fit to a 1∶1 Langmuir binding model, and oligomer and fibril data were fit to a heterogeneous ligand binding model. ΔRU: delta of the response units. t/s: time in seconds.
Table 1.
Overview of kinetic rates for scFv-IC16 binding to different Aβ(1–42) assembly states obtained within the single-cycle kinetic SPR experiments.
Figure 3.
Kinetic rates obtained for scFv-IC16 binding to different immobilized Aβ(1–42) assembly states.
Association rate constants (ka) were plotted against dissociation rate constants (kd). The dissociation constant (KD) can be extracted from the diagonal lines. Circles, squares and triangles correspond to data from interactions with monomers, oligomers and fibrils, respectively, whereas filled symbols represent data for the second binding site. All data was determined with the heterogeneous fitting model. The grey circle represents monomer data obtained with a 1∶1 binding model.