Thermodynamic analysis of lysozyme adsorbed to silica

Adsorption behavior of a human monoclonal antibody at hydrophilic and hydrophobic surfaces

“…This would be consistent with the known reduction in the rate of H/D exchange for lysozyme adsorbed to the silica surface compared with lysozyme in bulk solution. 33 While TIRF necessarily required very low concentrations to obey Leveque conditions, there is no such requirement for analysis of steady-state adsorption by NR; since the NR experiments were intended to capture the molecular nature of the adsorbed layers after ~30 min equilibration. The range of concentrations under which mAb-1 was adsorbed to surfaces represented concentrations that may reasonably be encountered during early formulation development.…”

“…We initially assumed a 70% exchange, in accordance with time course data for lysozyme adsorbed to silica (SiO 2 ) particles. 33 No calculation of H/D exchange based on a known hydrogen bonding pattern could be made since the 3-dimensional structure for mAb-1 has not been determined. Equation 3 was used to calculate the protein fraction of the layer covering a surface (a) from the fitted SLD (ρ) made up from contributions of the protein and subphase.…”

Measuring hydrogen–deuterium exchange in protein monolayers

“…[27] and references therein). Recently, techniques such as mass spectrometry [28] and nuclear magnetic resonance [29,30] have been used to describe conformational changes in proteins. While the solvent accessibility and HDX of adsorbed protein are most likely related to a variety of factors, our approach allows the HDX of both solution and surface samples to occur in identical environments, such as pH, which helps make comparisons of amide exposure relevant to structural information.…”

Interrogating protonated/deuterated fibronectin fragment layers adsorbed to titania by neutron reflectivity and their concomitant control over cell adhesion

“…Similarly, the surface fraction of the more stable mutant (FIII9 0 10-H2P) was 34% and 18% for the deuterated and protonated forms, respectively. All proteins showed a transition from monolayer to bilayer between 30 and 100 mg l 21 , with the protein longitudinal orientation moving away from the plane of the TiO 2 surface at high concentrations. Baby hamster kidney cells adherent to TiO 2 surfaces coated with the proteins (100 mg l…”

“…The histogrammed outputs for the corresponding data fits clearly show these distinct trends for the two isotopic forms during the wash step (electronic supplementary material, figure S7). When comparing the Monte Carlo generated histograms, it is interesting to note that the protein fraction of the lower and upper layers for either FIII9 0 10 or FIII9 0 10-H2P are very similar either at 200 mg l 21 or after the wash (electronic supplementary material, figures S6 and S7). This is not the case for the corresponding deuterated protein layers where the upper layer is largely washed off.…”