Adsorption of antibody therapeutics to air-liquid interfaces can enhance aggregation, particularly when the solution does not contain protective surfactant or when the surfactant is diluted as occurs during preparation of intravenous infusion bags. The ability to predict an antibody's propensity for interfacially mediated aggregation is particularly useful during product development to ensure the quality, potency, and safety of the therapeutic. To develop a predictive tool, we investigated the surface pressure and surface excess of a panel of 16 antibodies as well as determined their aggregation propensity at the air-liquid interface in an agitation stress model. Our data demonstrated that the initial rate of surface pressure increase upon antibody adsorption to the air-liquid interface strongly predicted the extent of agitation-induced aggregation. Other factors, including the hydrophobicity, equilibrium surface pressure, and interfacial concentration of an antibody, were not adequate predictors of its susceptibility to aggregation. In addition to developing a predictive tool, we extended the interfacial characterization to better understand the mechanisms of antibody aggregation at an air-liquid interface during agitation stress. We believe that the kinetics of antibody rearrangement and conformational change after adsorbing to the interface, leading to the development of attractive antibody-antibody interactions, dictated the extent of aggregation. Overall, our results demonstrate how surface pressure measurements can be implemented as a rapid screening tool for the identification of antibodies with a high propensity to aggregate upon adsorption to an air-liquid interface while also furthering our understanding of interfacially mediated protein aggregation.
To implement the molecular recognition properties of membrane proteins for applications including biosensors and
diagnostic arrays, the construction of a biomimetic platform capable of maintaining protein structure and function is
required. In this paper, we describe a tethered phospholipid vesicle assembly that overcomes the major limitations
of planar supported lipid bilayers and alternative biomimetic membrane platforms and characterize it using quartz
crystal microbalance with dissipation monitoring (QCM-D) and fluorescence microscopy. We provide evidence of
a one-step mechanism for bilayer formation and monitor the subsequent adsorption and binding of streptavidin,
vesicles, and streptavidin-coated microspheres. For all three species, we identify a critical surface density above which
a significant amount of coupled interstitial water contributes to the response of the quartz resonator in a phenomenon
similar to dynamic coupling due to surface roughness. A Sauerbrey-type analysis is sufficient to accurately interpret
the QCM-D results for streptavidin binding if water is treated as an additional inertial mass, but viscoelastic models
must be invoked for vesicle and microsphere binding. Additionally, we present evidence of vesicle flattening, possibly
enhanced by a biotin-mediated membrane−membrane interaction.
AMT helps in ocular surface reconstruction, promotes rapid epithelial healing and partially restores limbal stem cell function. It can be considered as an effective modality for the ocular surface restoration in chemical and thermal injuries in selected cases. Success rates in acute and chronic cases are comparable.
We show that Treg are increased in esophageal tissue of EoE subjects compared with GERD and HC subjects. The present study illustrates another possible mechanism involved in EoE that implicates impairment of immune homeostasis.
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