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The average hydrodynamic diameter, polydispersity, and zeta potential of PLGA NPs were characterized by dynamic light scattering and zeta potential measurements.
The NPs were dispersed in doubly distilled water, finally containing 1. The average diameter of NPs was — nm, and each NP contained approximately 4— carboxyl groups available for covalent binding of peptides. This attachment strategy resulted in uniformly oriented peptides of the two types. B-cell purification Peripheral blood mononuclear cells PBMCs were isolated from healthy donors rheumatoid arthritis pathogenesis pdf RA patients as described previously [ 38 ].
A higher purity of B cells is needed for the peptide-binding and for the cytotoxicity assays. Antibody secretion was determined by ELISA from the supernatant of in vitro stimulated B cells, with plates coated as already described.
The biotinylated βCit or βArg peptides were coupled to NeutrAvidin-labeled yellow—green microspheres 1 μm diameter; Thermo Fisher Scientific Inc. The peptide-coated fluorescent ízületi fájdalomkefe were added to B cells at fold excess and the samples were incubated for 1 hour at 4 °C. The spots were developed after 18 hours by biotinylated detection monoclonal antibodies, streptavidin HRP, and substrate Mabtech.
Pooled human serum or heat-inactivated serum was added at dilution for 1 hour. The bifunctional, fluorescein-containing NPs covered with the targeting βCit and the effector CNNQK peptides were added to B cells at fold excess, and then the samples were incubated for 1 hour on ice.
Statistical analysis For the statistical analysis of data, the Mann—Whitney test Fig.