n/a

The normal way of introducing the notion of derivative is by means of the limit of an incremental ratio that can assume three forms, depending the used translations as we saw in Chaps. 1 and 4. On the other hand, in those derivatives the limit operation is done over a set of points uniformly spaced: a linear scale was used. Here we present an alternative derivative, that is valid only for t {\ensuremath{>}} 0 or t {\ensuremath{<}} 0 and uses an exponential scale

Geobacter sulfurreducens is a sediment bacterium that contains a large number of multiheme cytochromes. The family of five c7 triheme periplasmic cytochromes from Geobacter sulfurreducens shows structural diversity of the heme core. Structural characterization of the relative orientation of the axial ligands of these proteins by 13C-paramagnetic NMR was carried out. The structures in solution were compared with those obtained by X-ray crystallography. For some hemes significant differences exist between the two methods such that orientation of the magnetic axes obtained from NMR data and the orientation taken from the X-ray coordinates differ. The results allowed the orientation of the magnetic axes to be defined confidently with respect to the heme frame in solution, a necessary step for the use of paramagnetic constraints to improve the complete solution structure of these proteins.

The phase transformations of the surfactant Triton X-100 were investigated by differential scanning calorimetry (DSC), polarized optical microscopy (POM), and dielectric relaxation spectroscopy (DRS). In particular, crystallization was induced at different cooling rates comprised between 13 and 0.5 K min–1. Vitrification was detected by both DSC and DRS techniques with a glass transition temperature of 212 K (measured on heating by DSC) allowing classifying Triton X-100 as a glass former. A fully amorphous material was obtained by cooling at a rate ≥10 K min–1, while crystallization was observed for lower cooling rates. The temperature of the onset of melt-crystallization was found to be dependent on the cooling scan rate, being higher the lower was the scan rate. In subsequent heating scans, the material undergoes cold-crystallization except if cooled previously at a rate ≤1 K min–1. None of the different thermal histories led to a 100% crystalline material because always the jump typical of the glass transformation in both heat flux (DSC) and real permittivity (DRS) is observed. It was also observed that the extent/morphology of the crystalline phase depends on the degree of undercooling, with higher spherulites developing for lower undercooling degree (24 K ≤ Tm – Tcr ≤ 44 K) in melt-crystallization and a grain-like morphology emerging for Tm – Tcr ≈ 57 K either in melt- or cold-crystallization. The isothermal cold- and melt-crystallizations were monitored near above the calorimetric glass transition temperature by POM (221 K) and real-time DRS (Tcr = 219, 220, and 221 K) to evaluate the phase transformation from an amorphous to a semicrystalline material. By DRS, the α-relaxation associated with the dynamic glass transition was followed, with the observation that it depletes upon both type of crystallizations with no significant changes either in shape or in location. Kinetic parameters were obtained from the time evolution of the normalized permittivity according to a modified Avrami model taking in account the induction time. The reason the isothermal crystallization occurs to a great extent in the vicinity of the glass transition was rationalized as the simultaneous effect of (i) a high dynamic fragile behavior and (ii) the occurrence of catastrophic nucleation/crystal growth probably enabled by a preordering tendency of the surfactant molecules. This is compatible with the estimated low Avrami exponent (1.12 ≤ n ≤ 1.6), suggesting that relative short length scale motions govern the crystal growth in Triton X-100 coherent with the observation of a grainy crystallization by POM.

In this work, we systematically evaluated the potential of using boronic acid functionalized magnetic particles in the capturing of human immunoglobulin G under typical mammalian cell culture conditions. For comparison, Protein A coated magnetic particles were also used. The binding pH was found to significantly influence the adsorption isotherms of boronic acid particles with the higher capacities (0.216 g IgG/g support) being observed at pH 7.4. Comparatively, this value was 0.109 g IgG/g support, for Protein A particles under the same conditions. Both particles revealed very fast adsorption kinetics with more than 70% of the maximum binding capacity being achieved in a few seconds. The effect of glucose and lactate, which are known to interact with boronic acid, was evaluated. For glucose, the binding capacity was significantly influenced by the pH and decreased as pH increased. At pH 9.5, a 70% lower binding capacity was observed for glucose concentrations as low as 0.5 g/l. The effect of lactate was less pronounced and almost pH independent reaching at most 20% decrease in binding capacity. Nevertheless, the effect of both molecules was always lower at pH 7.4. The optimization of the elution conditions enabled complete recovery of bound IgG from boronic acid particles using 50mM Tris-HCl, 200 mM sorbitol, 200 mM NaCl at pH 8.5.