Suresh K. Bhatia

Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces

A procedure for covalent immobilization of functional proteins on silica substrates was developed using thiol-terminal silanes and heterobifunctional cross-linkers. Using this procedure, a high density of functional antibodies was bound to glass cover slips and silica fibers. The amount of anti-IgG antibody immobilized was determined to be in the range of 0.66 to 0.96 ng/mm2 using radiolabeled antibody. The relative amount of IgG antigen bound by the immobilized antibody (0.37 to 0.55 mol antigen/mol antibody) was three to five times greater than other investigators have reported. In addition, the amount of protein nonspecifically adsorbed to the antibody-coated surface was further reduced by the addition of blocking agents so that nonspecific adsorption of protein antigens represented only 2-6% of the total antigen binding. With this low background, IgG antigen binding could be measured at levels as low as 150 fmol when an antigen concentration of 3 pmol/ml was applied. The process for antibody immobilization is straightforward, easy to perform, and adaptable for modifying mass quantities of biosensor components.

Variation of the pore structure of coal chars during gasification

The variation of the pore structure of several coal chars during gasification in air and carbon dioxide was studied by argon adsorption at 87 K and CO2 adsorption at 273 K. It is found that the surface area and volume of the small pores ( 10 A for air gasification is constant over a wide range of conversion (>20%), while for CO2 gasification similar results are obtained using the total surface area. However, in the early stages of gasification (<20%) the normalized reaction rate is much higher than that in the later stage of gasification, due to existence of more inaccessible pores in the beginning of gasification. The inaccessibility of the micropores to adsorption at low and ambient temperatures is confirmed by the measurement of the helium density of the coal chars. The random pore model can fit the experimental data well and the fitted structural parameters match those obtained by physical gas adsorption for coal chars without closed pores.

Antibody immobilization using heterobifunctional crosslinkers

Covalent attachment of functional proteins to a solid support is important for biosensors. One method employs thiol-terminal silanes and heterobifunctional crosslinkers such as N-succinimidyl 4-maleimidobutyrate (GMBS) to immobilize proteins through amino groups onto glass, silica, silicon or platinum surfaces. In this report, several heterobifunctional crosslinkers are compared to GMBS for their ability to immobilize active antibodies onto glass cover slips at a high density. Antibodies were immobilized at densities of 74-220 ng/cm2 with high levels of specific antigen binding. Carbohydrate-reactive crosslinkers were also compared to GMBS using a fiber optic biosensor to detect fluorescently-labeled antigen. At the concentrations tested, the antibodies immobilized with carbohydrate-reactive crosslinkers bound more antigen than GMBS immobilized antibodies as indicated by the fluorescence signal.

Structural ordering of coal char during heat treatment and its impact on reactivity

The effect of heat treatment on the structure of an Australian semi-anthracite char was studied in detail in the 850-1150degreesC temperature range using XRD, HRTEM, and electrical resistivity techniques. It was found that the carbon crystallite size in the char does not change significantly during heat treatment in the temperature range studied, for both the raw coal and its ash-free derivative obtained by acid treatment. However, the fraction of the organized carbon in the raw coal chars, determined by XRD, increased with increase of heat treatment time and temperature, while that for the ash-free coal chars remained almost unchanged. This suggests the occurrence of catalytic ordering during heat treatment, supported by the observation that the electrical resistivity of the raw coal chars decreased with heat treatment, while that of the ash-free coal chars did not vary significantly. Further confirmatory evidence was provided by high resolution transmission electron micrographs depicting well-organized carbon layers surrounding iron particles. It is also found that the fraction of organized carbon does not reach unity, but attains an apparent equilibrium value that increases with increase in temperature, providing an apparent heat of ordering of 71.7 kJ mol(-1) in the temperature range studied. Good temperature-independent correlation was found between the electrical resistivity and the organized carbon fraction, indicating that electrical resistivity is indeed structure sensitive. Good correlation was also found between the electrical resistivity and the reactivity of coal char. All these results strongly suggest that the thermal deactivation is the result of a crystallite-perfecting process, which is effectively catalyzed by the inorganic matter in the coal char. Based on kinetic interpretation of the data it is concluded that the process is diffusion controlled, most likely involving transport of iron in the inter-crystallite nanospaces in the temperature range studied. The activation energy of this transport process is found to be very low, at about 11.8 kJ mol(-1), which is corroborated by model-free correlation of the temporal variation of organized carbon fraction as well as electrical resistivity data using the superposition method, and is suggestive of surface transport of iron

Molecular transport in nanopores: a theoretical perspective

Molecular transport in nanopores plays a central role in many emerging nanotechnologies for gas separation and storage, as well as in nanofluidics. Theories of the transport provide an understanding of the mechanisms that influence the transport and their interplay, and can lead to tractable models that can be used to advance these nanotechnologies through process analysis and optimisation. We review some of the most influential theories of fluid transport in small pores and confined spaces. Starting from the century old Knudsen formulation, the dusty gas model and several other related approaches that share a common point of departure in the Maxwell-Stefan diffusion equations are discussed. In particular, the conceptual basis of the models and the validity of the assumptions and simplifications necessary to obtain their final results are analysed. It is shown that the effect of adsorption is frequently either neglected, or treated on an ad hoc basis, such as through the division of the pore flux into gas-phase and surface diffusion contributions. Furthermore, while it is commonplace to assume that cross-sectional pressure is uniform, it is demonstrated that this violates the Gibbs-Duhem relation and that it is the chemical potential that essentially remains constant in the cross-section, as near-equilibrium density profiles are preserved even during transport. The Dusty Gas model and Maxwell-Stefan model for surface diffusion are analysed, and their strengths and weaknesses discussed, illustrating the use of conflicting choices of frames of reference in the former case, and the importance of assigning appropriate values for the binary diffusivity in the latter case. The oscillator model, developed in this laboratory, which is exact in the low density limit under diffuse reflection conditions, is shown to represent an advance on the classical Knudsen formula, although the latter frequently appears as a fundamental part of many transport models. The distributed friction model, also developed in this laboratory for the study of multi-component transport at any Knudsen number is discussed and compared with previous approaches. Finally, the outlook for theory and future research needs are discussed.

Rational pattern design for in vitro cellular networks using surface photochemistry

The ability to create patterns of specific silane monolayers by deep ultraviolet lithography has been previously demonstrated, and preliminary attempts have been made to use these patterns to control adhesion and outgrowth of neurons and other types of mammalian cells. Here we report characterization of the mechanisms involved in these photoinitiated processes and their utility in various strategies for creating patterns for biologically relevant systems. We have divided the mechanisms into three general classes. The first is surface photolysis of the silane monolayer, which appears to proceed by a purely photochemical mechanism. The second mechanism involves direct photochemical conversion of a terminal functional group on a silane monolayer into a species with altered properties, e.g., the conversion of a thiol to a more oxidized form that inhibits the subsequent adhesion of proteins. The third is a photolytic degradation of the monolayer. The mechanisms have been probed by x‐ray photoelectron spectrosc...

New Method for Atomistic Modeling of the Microstructure of Activated Carbons Using Hybrid Reverse Monte Carlo Simulation

We propose a new hybrid reverse Monte Carlo (HRMC) procedure for atomistic modeling of the microstructure of activated carbons whereby the guessed configuration for the HRMC construction simulation is generated using the characterization results (pore size and pore wall thickness distributions) obtained by the interpretation of argon adsorption at 87 K using our improved version of the slit-pore model, termed the finite wall thickness (FWT) model (Nguyen, T. X.; Bhatia, S. K. Langmuir 2004, 20, 3532) . This procedure overcomes limitations arising from the use of short-range potentials in the conventional HRMC method, which make the latter unsuitable for carbons such as activated carbon fibers that are anisotropic with medium-range ordering induced by their complex pore structure. The newly proposed approach is applied specifically for the atomistic construction of an activated carbon fiber ACF15, provided by Kynol Corporation (Nguyen, T. X.; Bhatia, S. K. Carbon 2005, 43, 775) . It is found that the PSD of the ACF15s constructed microstructure is in good agreement with that determined using argon adsorption at 87 K. Furthermore, we have also found that the use of the Lennard-Jones (LJ) carbon-fluid interaction well depth obtained from scaling the flat graphite surface-fluid interaction well depth taken from Steele (Steele, W. A. Surf. Sci. 1973, 36, 317) provides an excellent prediction of experimental adsorption data including the differential heat of adsorption of simple gases (Ar, N(2), CH(4), CO(2)) over a wide range of temperatures and pressures. This finding is in agreement with the enhancement of the LJ carbon-fluid well depth due to the curvature of the carbon surface, found by the use of ab initio calculations (Klauda, J. B.; Jiang, J.; Sandler, S. I. J. Phys. Chem. B 2004, 108, 9842) .

AXIAL SEGREGATION OF PARTICLES IN A HORIZONTAL ROTATING CYLINDER

We report on an experimental study of the axial segregation process for particle mixture of different sizes. The effect of particle volume fraction and rotational speed on band formation is considered. A mechanism for the segregation process is presented based on which criteria for axial segregation are discussed

Tractable molecular theory of transport of Lennard-Jones fluids in nanopores

We present here a tractable theory of transport of simple fluids in cylindrical nanopores, which is applicable over a wide range of densities and pore sizes. In the Henry law low-density region the theory considers the trajectories of molecules oscillating between diffuse wall collisions, while at higher densities beyond this region the contribution from viscous flow becomes significant and is included through our recent approach utilizing a local average density model. The model is validated by means of equilibrium as well nonequilibrium molecular dynamics simulations of supercritical methane transport in cylindrical silica pores over a wide range of temperature, density, and pore size. The model for the Henry law region is exact and found to yield an excellent match with simulations at all conditions, including the single-file region of very small pore size where it is shown to provide the density-independent collective transport coefficient. It is also shown that in the absence of dispersive interactions the model reduces to the classical Knudsen result, but in the presence of such interactions the latter model drastically overpredicts the transport coefficient. For larger micropores beyond the single-file region the transport coefficient is reduced at high density because of intermolecular interactions and hindrance to particle crossings leading to a large decrease in surface slip that is not well represented by the model. However, for mesopores the transport coefficient increases monotonically with density, over the range studied, and is very well predicted by the theory, though at very high density the contribution from surface slip is slightly overpredicted. It is also seen that the concept of activated diffusion, commonly associated with diffusion in small pores, is fundamentally invalid for smooth pores, and the apparent activation energy is not simply related to the minimum pore potential or the adsorption energy as generally assumed.

Comparisons of diffusive and viscous contributions to transport coefficients of light gases in single-walled carbon nanotubes

We examine here the relative importance of different contributions to transport of light gases in single walled carbon nanotubes, using methane and hydrogen as examples. Transport coefficients at 298 K are determined using molecular dynamics simulation with atomistic models of the nanotube wall, from which the diffusive and viscous contributions are resolved using a recent approach that provides an explicit expression for the latter. We also exploit an exact theory for the transport of Lennard-Jones fluids at low density considering diffuse reflection at the tube wall, thereby permitting the estimation of Maxwell coefficients for the wall reflection. It is found that reflection from the carbon nanotube wall is nearly specular, as a result of which slip flow dominates, and the viscous contribution is small in comparison, even for a tube as large as 8.1 nm in diameter. The reflection coefficient for hydrogen is 3–6 times as large as that for methane in tubes of 1.36 nm diameter, indicating less specular reflection for hydrogen and greater sensitivity to atomic detail of the surface. This reconciles results showing that transport coefficients for hydrogen and methane, obtained in simulation, are comparable in tubes of this size. With increase in adsorbate density, the reflection coefficient increases, suggesting that adsorbate interactions near the wall serve to roughen the local potential energy landscape perceived by fluid molecules.