Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages.

Nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118-131 and 137-140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation.

Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages

Nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118-131 and 137-140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation. © 2013 Elsevier Ltd.

Design consideration of a circular type magnetic flux pump device

As a variation of the thermally actuated flux pump and the linear type magnetic flux pump (LTMFP), the circular type magnetic flux pump (CTMFP) device is proposed to magnetize a circular shape type-II superconducting thin film and bulk. The basic concept is the same as the thermally actuated flux pump: a circularly symmetric traveling magnetic field is generated below a circular shape superconductor to increase its trapping field. However, this traveling field is created by the three phase windings instead of heating gadolinium block. Apart from the LTMFP, the three phase windings are wound concentrically instead of linearly. The speed of the traveling field is controlled by the AC frequency and the magnitude of the field is controlled by the magnitudes of AC currents. In addition, a coil with DC current is wound around the three phase windings to provide a background field. The concept design is presented in this paper. The magnetic waveforms are analysed numerically by the COMSOL 3.5a software. The impedances of the three phase windings are calculated and a corresponding circuit design is presented. This rig can be used as an advanced tool to study the flux pump behavior of a circular shape superconductor. © 2002-2011 IEEE.

The investigation of sweeping speed on the magnetization of a 2 inches diameter YBa2Cu3O7-δ thin film with the circular-type magnetic flux pump

A circular-type magnetic flux pump (CTMFP) device was built to study the flux dynamics on a 2-inch-diameter YBCO thin film. This CTMFP is composed of two CTMFP coils, with each CTMFP coil containing concentric three-phase windings and a dc winding. We connected the three-phase windings to the output of a commercial inverter. By changing the output frequency of the inverter, the sweeping speed of the circular-shaped travelling magnetic wave can be changed. The connection of the phase coils follows the forward consequence, so that the circular-shaped travelling magnetic wave travels inward to the center. The output frequency f was changed from f = 0.01 to 1000.0 Hz. The YBCO sample was sandwiched between the two CTMFP coils to experience the circular-shaped travelling magnetic wave. It was found that the increase of the flux density in the center of the film is independent of the sweeping frequency. In high frequency f = 1000.0Hz, even if the waveform had changed a lot, the increment is still the same as in low frequencies. © 2012 IEEE.

Magnetic switching and anisotropy in nanomagnets: the influence of size and shape

Arrays of nanomagnets were fabricated out of Ni80Fe14Mo5 in the lateral size range 500-30nm and the thickness range 3-20nm. Elliptical, triangular, square, pentagonal and circular geometries were all considered. The magnetic properties of these nanomagnets were probed rapidly and non-invasively using a high sensitivity magneto-optical method.

Controlling magnetic ordering in coupled nanomagnet arrays

We have fabricated using high-resolution electron beam lithography circular magnetic particles (nanomagnets) of diameter 60 nm and thickness 7 nm out of the common magnetic alloy supermalloy. The nanomagnets were arranged on rectangular lattices of different periods. A high-sensitivity magneto-optical method was used to measure the magnetic properties of each lattice. We show experimentally how the magnetic properties of a lattice of nanomagnets can be profoundly changed by the magnetostatic interactions between nanomagnets within the lattice. We find that simply reducing the lattice spacing in one direction from 180 nm down to 80 nm (leaving a gap of only 20 nm between edges) causes the lattice to change from a magnetically disordered state to an ordered state. The change in state is accompanied by a peak in the magnetic susceptibility. We show that this is analogous to the paramagnetic-ferromagnetic phase transition which occurs in conventional magnetic materials, although low-dimensionality and kinetic effects must also be considered.

Vortex migration caused by travelling magnetic wave in a 2 in. diameter YBa2Cu3O7-δ thin film

We provide experimental evidence for a vortex migration phenomenon in YBa2Cu3O7-δ (YBCO) thin film caused by travelling magnetic wave. The experiment is carried out on a 2 in. diameter YBCO thin film with a circular-type magnetic flux pump. We found that the travelling wave helps the vortices migrate into the centre of the sample: after the zero-field cooling process, the increase of the flux density in the centre is four times larger than the amplitude of the travelling wave. The reason for this massive vortex migration is probably due to the magnetic stress variation caused by the travelling wave: the magnetic stress increases locally in the crest region while decreases locally in the trough region, which could help the vortices to move locally. A comparison shows that the magnetization by standing wave can be easily predicted by Bean's model while travelling wave causes vortex migration generally much larger than the prediction of Bean's model. It is possible that travelling magnetic wave can be an effective way to magnetize a type II superconductor in considering this unusual vortex dynamics. © 2013 AIP Publishing LLC.