Henrich Frielinghaus

Self-assembly and gelation properties of α-helix versus β-sheet forming peptides

We have investigated the self-assembly and gelation properties of a set of four octa-peptides: AEAEAKAK, AEAKAEAK, FEFEFKFK and FEFKFEFK. The phenylalanine based peptides adopt β-sheet conformations in solution and the alanine based peptides form α-helices. No self-assembly in solution was observed for AEAKAEAK but AEAEAKAK was found to self-assemble forming thick, rigid fibres with a diameter of ∼6 nm. These fibres were composed of two fibrils aggregating side by side to form “pearl-necklace” morphologies. No gelation was observed for AEAEAKAK in the concentration range investigated (0 to 100 mg ml−1). In contrast, both phenylalanine based peptides were found to self-assemble in solution and to form hydrogels at an initial concentration of ∼8 mg ml−1. Similar morphologies were observed for both peptides corresponding to a relatively homogeneous dense network of semi-flexible fibres with a mesh size of ∼15 to 30 nm depending on the concentration. The fibre diameter was found to be ∼4 nm in good agreement with models found in the literature. TEM micrographs clearly showed that these fibres have a helicoidal or twisted structure. Comparison of TEM with AFM data highlighted the influence of substrate chemistry on the macromolecular assembly of small peptides. In contrast small angle neutron scattering (SANS) approaches, which allow for the probing of hydrogel morphology and structure without the need for sample preparation on solid substrates, provide vital data on hydrogel morphology in solution.

Hydration and interactions in protein solutions containing concentrated electrolytes studied by small-angle scattering

During protein crystallization and purification, proteins are commonly found in concentrated salt solutions. The exact interplay of the hydration shell, the salt ions, and protein-protein interactions under these conditions is far from being understood on a fundamental level, despite the obvious practical relevance. We have studied a model globular protein (bovine serum albumin, BSA) in concentrated salt solutions by small-angle neutron scattering (SANS). The data are also compared to previous studies using SAXS. The SANS results for dilute protein solutions give an averaged volume of BSA of 91,700 Å(3), which is about 37% smaller than that determined by SAXS. The difference in volume corresponds to the contribution of a hydration shell with a hydration level of 0.30 g g(-1) protein. The forward intensity I(0) determined from Guinier analysis is used to determine the second virial coefficient, A(2), which describes the overall protein interactions in solution. It is found that A(2) follows the reverse order of the Hofmeister series, i.e. (NH(4))(2)SO(4) < Na(2)SO(4) < NaOAc < NaCl < NaNO(3) < NaSCN. The dimensionless second virial coefficient B(2), corrected for the particle volume and molecular weight, has been calculated using different approaches, and shows that B(2) with corrections for hydration and the non-spherical shape of the protein describes the interactions better than those determined from the bare protein. SANS data are further analyzed in the full q-range using liquid theoretical approaches, which gives results consistent with the A(2) analysis and the experimental structure factor.

Origin of Buckling Phenomenon during Drying of Micrometer-Sized Colloidal Droplets

The origin of the buckling of micrometer-sized colloidal droplets during evaporation-induced self-assembly (EISA) has been elucidated using electron microscopy and small-angle neutron scattering. Doughnut-like assembled grains with varying aspect ratios are formed during EISA at different physicochemical conditions. It has been revealed that this phenomenon is better explained by an existing hypothesis based on the formation of a viscoelastic shell of nanoparticles during drying than by other existing hypotheses based on the inertial instability of the initial droplets and hydrodynamic instability due to thermocapillary forces. This conclusion was further supported by the arrest of buckling through modification of the colloidal interaction in the initial dispersion.

Preferential hydration of lysozyme in water/glycerol mixtures: A small-angle neutron scattering study

In solution small-angle neutron scattering has been used to study the solvation properties of lysozyme dissolved in water/glycerol mixtures. To detect the characteristics of the protein-solvent interface, 35 different experimental conditions (i.e., protein concentration, water/glycerol fraction in the solvent, content of deuterated compounds) have been considered and a suitable software has been developed to fit simultaneously the whole set of scattering data. The average composition of the solvent in the close vicinity of the protein surface at each experimental condition has been derived. In all the investigated conditions, glycerol resulted especially excluded from the protein surface, confirming that lysozyme is preferentially hydrated. By considering a thermodynamic hydration model based on an equilibrium exchange between water and glycerol from the solvation layer to the bulk, the preferential binding coefficient and the excess solvation number have been estimated. Results were compared with data previously derived for ribonuclease A in the same mixed solvent: even if the investigated solvent compositions were very different, the agreement between data is noticeable, suggesting that a unique mechanism presides over the preferential hydration process. Moreover, the curve describing the excess solvation number as a function of the solvent composition shows the occurrence of a region of maximal hydration, which probably accounts for the changes in protein stability detected in the presence of cosolvents.

Adsorption of supercritical CO2 in aerogels as studied by small-angle neutron scattering and neutron transmission techniques.

Small-angle neutron scattering (SANS) has been used to study the adsorption behavior of supercritical carbon dioxide (CO2) in porous Vycor glass and silica aerogels. Measurements were performed along two isotherms (T=35 and 80 degrees C) as a function of pressure (P) ranging from atmospheric up to 25 MPa, which corresponds to the bulk fluid densities ranging from rho(CO2) approximately 0 to 0.9 gcm3. The intensity of scattering from CO2-saturated Vycor porous glass can be described by a two-phase model which suggests that CO2 does not adsorb on the pore walls and fills the pore space uniformly. In CO2-saturated aerogels an adsorbed phase is formed with a density substantially higher that of the bulk fluid, and neutron transmission data were used to monitor the excess adsorption at different pressures. The results indicate that adsorption of CO2 is significantly stronger in aerogels than in activated carbons, zeolites, and xerogels due to the extremely high porosity and optimum pore size of these materials. SANS data revealed the existence of a compressed adsorbed phase with the average density approximately 1.07 gcm3, close to the density corresponding to closely packed van der Waals volume of CO2. A three-phase model [W. L. Wu, Polymer 23, 1907 (1982)] was used to estimate the volume fraction phi3 of the adsorbed phase as a function of the fluid density, and gave phi3 approximately 0.78 in the maximum adsorption regime around rho(CO2) approximately 0.374 gcm3. The results presented in this work demonstrate the utility of SANS combined with the transmission measurements to study the adsorption of supercritical fluids in porous materials.

KWS-2, the high intensity / wide Q-range small-angle neutron diffractometer for soft-matter and biology at FRM II

The KWS-2 small-angle neutron diffractometer operated by JCNS at FRM II is upgraded and optimized towards high intensity and wide Q-range studies of mesoscopic structures and structural changes due to rapid kinetics and becomes a dedicated SANS instrument to soft-matter and biology. The high intensity permits fast measurement of small or weak scattering samples and time resolved structural studies with a time resolution up to 100ms. The possibility to cover up to four decades in Q will soon enable structural investigation over a wide length scale, between several A and lμm. The characteristics and performance of the instrument in the conventional pinhole mode is detailed presented and the new upgrades currently in progress and aiming for boosting the instrument performance towards higher intensity and wider Q-range are reported.

KWS-1 high-resolution small-angle neutron scattering instrument at JCNS: current state

The KWS-1 small-angle neutron scattering (SANS) instrument operated by the Julich Centre for Neutron Science (JCNS) at the research reactor FRM II of the Heinz Maier-Leibnitz Zentrum in Garching near Munich has been recently upgraded. The KWS-1 instrument was updated, from its active collimation apertures to the detector cabling. Most of the parts of the instrument were installed for the first time, including a broadband polarizer, a large-cross-section radio-frequency spin flipper, a chopper and neutron lenses. A custom-designed hexapod in the sample position allows heavy loads and precise sample positioning in the beam for conventional SANS experiments as well as for grazing-incidence SANS under applied magnetic field. With the foreseen in situ polarization analysis the main scientific topic of the instrument tends towards magnetism. The performance of the polarizer and flipper was checked with a polarized 3He cell at the sample position. The results of these checks and a comparison of test measurements on a ferrofluid in a magnetic field with polarized and nonpolarized neutrons are presented.

Thermal composition fluctuations near the isotropic Lifshitz critical point in a ternary mixture of a homopolymer blend and diblock copolymer

We have studied thermal composition fluctuations of a ternary symmetric homopolymer/diblock copolymer system of PEE/PDMS/PEE-PDMS [PEE and PDMS being poly(ethyl ethylene) and poly(dimethyl siloxane), respectively] in its disordered state with small angle neutron scattering for concentration Φ of diblocks up to 15%. The phase diagram shows three characteristic regimes; (1) below the Lifshitz concentration ΦLL≅9%; (2) in the very near vicinity of the Lifshitz concentration; and (3) above ΦLL. In the regime (1) of low diblock content the maximum neutron intensity is obtained at Q=0 and phase separation into macroscopic large domains is observed at low temperatures. With increasing diblock content the thermal fluctuations indicate a crossover from 3d-Ising to isotropic Lifshitz critical behavior with critical exponents of the susceptibility γ=(1.62±0.01) and correlation length ν=(0.99±0.04) appreciably larger than in the 3d-Ising case. In the structure factor this crossover is accompanied by a strong reductio...

Reinforcement of model filled elastomers: synthesis and characterization of the dispersion state by SANS measurements

This work is the first part of a study devoted to the understanding and the determination of the molecular mechanisms that are at the origin of the specific properties shown by reinforced elastomers. Different model filled elastomers composed of cross-linked polyethylacrylate chains reinforced with grafted silica nanoparticles were prepared varying the reactivity of the coupling agent with the ethylacrylate monomers. They were synthetized applying and adapting the method developed by Ford et al. [11] which consists to polymerize a colloidal suspension of grafted silica particles in acrylate monomers. In this paper we will present how filled elastomers having different dispersion states can be prepared whilst keeping the same interactions between the particles and the polymer chains. The dispersion states were characterized by Small Angle Neutron Scattering. We found that there are two opposite effects which control the final dispersion state of these filled elastomers during the polymerization. The first one is a depletion mechanism favoring the formation of aggregates. The second one is a repulsive steric interaction due to the growth of polymer chains from the particle surfaces avoiding contacts between the silica inclusions. Using these results we can prepare sets of samples having the same particle/matrix interface but different dispersions states. By comparing their mechanical properties we should to able to estimate the relative weight of the dispersion state quality and the one of the particle/matrix interface on the mechanical behavior of these filled elastomers.

Dynamic properties of microemulsions modified with homopolymers and diblock copolymers: The determination of bending moduli and renormalization effects

The properties of bicontinuous microemulsions, consisting of water, oil, and a surfactant, can be modified by the addition of diblock copolymers (boosting effect) and homopolymers (inverse boosting effect) or a combination of both. Here, the influence of the addition of homopolymers (PEP(X) and PEO(X), X=5k or 10k molecular weight) on the dynamics of the surfactant layer is studied with neutron spin echo spectroscopy (NSE). Combining the results with the previous findings for diblock copolymers allows for a better separation of viscosity and bending modulus effects. With the addition of homopolymers, a significant increase of the relaxation rate compared to the pure microemulsion has been observed. The influence on the bending rigidity kappa is measured with NSE experiments. Homopolymer addition reduces kappa by up to Deltakappa approximately -0.5k(B)T, whereas the diblock copolymer yields an increase of kappa by approximately 0.3k(B)T. Comparison of the bending moduli that are obtained by analysis of the dynamics to those obtained from small angle neutron scattering (SANS) sheds light on the different renormalization length scales for NSE and SANS. Variation of the surfactant concentration at otherwise constant conditions of homopolymer or diblock-copolymer concentration shows that NSE results are leading to the pure bending rigidity, while the renormalized one is measured with SANS.