Sumit Mehan

Small-angle neutron scattering study of structure and interaction of nanoparticle, protein, and surfactant complexes.

Small-angle neutron scattering (SANS) measurements have been carried out from the multicomponent system composed of Ludox HS40 silica nanoparticle, bovine serum albumin (BSA) protein, and sodium dodecyl sulfate (SDS) surfactant in an aqueous system under the solution condition that all the components are negatively charged. Although the components are similarly charged, strong structural evolutions among them have been observed. The complexes of different components in pairs (nanoparticle-protein, nanoparticle-surfactant, and protein-surfactant) have been examined to correlate the role of each component in the three-component nanoparticle-protein-surfactant system. The nanoparticle-protein system shows depletion interaction induced aggregation of nanoparticles in the presence of protein. Both nanoparticle and surfactant coexist individually in a nanoparticle-surfactant system. In the case of a protein-surfactant system, the cooperative binding of surfactant with protein leads to micelle-like clusters of surfactant formed along the unfolded protein chain. The structure of the three-component (nanoparticle-protein-surfactant) system is found to be governed by the synergetic effect of nanoparticle-protein and protein-surfactant interactions. The nanoparticle aggregates coexist with the structures of protein-surfactant complex in the three-component system. The nanoparticle aggregation as well as unfolding of protein is enhanced in this system as compared to the corresponding two-component systems.

Cationic versus anionic surfactant in tuning the structure and interaction of nanoparticle, protein, and surfactant complexes.

The structure and interaction in complexes of anionic Ludox HS40 silica nanoparticle, anionic bovine serum albumin (BSA) protein, and cationic dodecyl trimethylammonium bromide (DTAB) surfactant have been studied using small-angle neutron scattering (SANS). The results are compared with similar complexes having anionic sodium dodecyl sulfate (SDS) surfactant (Mehan, S; Chinchalikar, A. J.; Kumar, S.; Aswal, V. K.; Schweins, R. Langmuir 2013, 29, 11290). In both cases (DTAB and SDS), the structure in nanoparticle-protein-surfactant complexes is predominantly determined by the interactions of the individual two-component systems. The nanoparticle-surfactant (mediated through protein-surfactant complex) and protein-surfactant interactions for DTAB, but nanoparticle-protein (mediated through protein-surfactant complex) and protein-surfactant interactions for SDS, are found to be responsible for the resultant structure of nanoparticle-protein-surfactant complexes. Irrespective of the charge on the surfactant, the cooperative binding of surfactant with protein leads to micellelike clusters of surfactant formed along the unfolded protein chain. The adsorption of these protein-surfactant complexes for DTAB on oppositely charged nanoparticles gives rise to the protein-surfactant complex-mediated aggregation of nanoparticles (similar to that of DTAB surfactant). It is unlike that of depletion-induced aggregation of nanoparticles with nonadsorption of protein-surfactant complexes for SDS in similarly charged nanoparticle systems (similar to that of protein alone). The modifications in nanoparticle aggregation as well as unfolding of protein in these systems as compared to the corresponding two-component systems have also been examined by selectively contrast matching the constituents.

Tuning of protein-surfactant interaction to modify the resultant structure.

Small-angle neutron scattering and dynamic light scattering studies have been carried out to examine the interaction of bovine serum albumin (BSA) protein with different surfactants under varying solution conditions. We show that the interaction of anionic BSA protein (pH7) with surfactant and the resultant structure are strongly modified by the charge head group of the surfactant, ionic strength of the solution, and mixed surfactants. The protein-surfactant interaction is maximum when two components are oppositely charged, followed by components being similarly charged through the site-specific binding, and no interaction in the case of a nonionic surfactant. This interaction of protein with ionic surfactants is characterized by the fractal structure representing a bead-necklace structure of micellelike clusters adsorbed along the unfolded protein chain. The interaction is enhanced with ionic strength only in the case of site-specific binding of an anionic surfactant with an anionic protein, whereas it is almost unchanged for other complexes of cationic and nonionic surfactants with anionic proteins. Interestingly, the interaction of BSA protein with ionic surfactants is significantly suppressed in the presence of nonionic surfactant. These results with mixed surfactants thus can be used to fold back the unfolded protein as well as to prevent surfactant-induced protein unfolding. For different solution conditions, the results are interpreted in terms of a change in fractal dimension, the overall size of the protein-surfactant complex, and the number of micelles attached to the protein. The interplay of electrostatic and hydrophobic interactions is found to govern the resultant structure of complexes.

Probing structural evolution in heat induced protein gelation by scattering techniques

The heating of a globular protein is known to elicit conformational changes in the protein molecules, resulting in the formation of a gel depending on the solution conditions. We have used dynamic light scattering (DLS) and small-angle neutron scattering (SANS) to investigate the phase behavior and structure evolution in heat induced gelation of Bovine Serum Albumin (BSA) protein as a function of pH and ionic strength. The gelation temperature is found to be increasing with increase in pH and decrease in ionic strength. The structure of the protein molecule remains intact very close to the gelation temperature. However, on further increase in temperature, the protein molecules form small aggregates which eventually lead to a network gel at gelation temperature. The gel structure is characterized by a mass fractal having a fractal dimension about 2.

Observation of dynamic equilibrium cluster phase in nanoparticle-polymer system

Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) have been used to investigate the existence of a cluster phase in a nanoparticle-polymer system. The nanoparticle-polymer system shows an interesting reentrant phase behavior where the charge stabilized silica nanoparticles undergo particle clustering and back to individual nanoparticles as a function of polymer concentration. This kind of phase behavior is believed to be directed by opposing attractive and repulsive interactions present in the system. The phase behavior shows two narrow regions of polymer concentration immediately before and after the two-phase formation indicating the possibility of the existence of some equilibrium clusters. DLS results show a much higher size of particles than individuals in these two regions which remains unchanged even after dilution. The SANS data show the evolution of attraction with increased volume fraction of the particles supporting the dynamic nature of these clusters.

Structural study of surfactant-dependent interaction with protein

Small-angle neutron scattering (SANS) has been used to study the complex structure of anionic BSA protein with three different (cationic DTAB, anionic SDS and non-ionic C12E10) surfactants. These systems form very different surfactant-dependent complexes. We show that the structure of protein-surfactant complex is initiated by the site-specific electrostatic interaction between the components, followed by the hydrophobic interaction at high surfactant concentrations. It is also found that hydrophobic interaction is preferred over the electrostatic interaction in deciding the resultant structure of protein-surfactant complexes.

Observation of two different fractal structures in nanoparticle, protein and surfactant complexes

Small angle neutron scattering has been carried out from a complex of nanoparticle, protein and surfactant. Although all the components are similarly (anionic) charged, we have observed strong interactions in their complex formation. It is characterized by the coexistence of two different mass fractal structures. The first fractal structure is originated from the protein and surfactant interaction and second from the depletion effect of first fractal structure leading the nanoparticle aggregation. The fractal structure of protein-surfactant complex represents to bead necklace structure of micelle-like clusters of surfactant formed along the unfolded protein chain. Its fractal dimension depends on the surfactant to protein ratio (r) and decreases with the increase in r. However, fractal dimension of nanoparticle aggregates in nanoparticle-protein complex is found to be independent of protein concentration and governed by the diffusion limited aggregation like morphology.