Physicochemical properties of immunolatex, prepared by incubation of negatively charged polystyrene microparticles with polyclonal rabbit IgGs, were determined by a variety of experimental techniques. These comprised dynamic light scattering (DLS), laser Doppler velocimetry (LDV) and atomic force microscopy (AFM). The particle diffusion coefficient, the hydrodynamic diameter, the electrophoretic mobility, the zeta potential and the suspension stability were determined as a function of pH for different ionic strengths. The deposition of the immunolatex on bare and polyallylamine (PAH) functionalized mica was investigated using the microfluidic oblique impinging-jet cell, with an in situ, real-time image analysis module. The particle deposition kinetics was acquired by a direct particle enumeration procedure. The measurements enabled us to determine the range of pH where the specific deposition of the immunolatex on these substrates was absent. We argue that the obtained results have practical significance for conducting efficient flow immunoassays governed by specific antigen/antibody interactions.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9820969PMC
http://dx.doi.org/10.3390/ijms24010692DOI Listing

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Physicochemical properties of immunolatex, prepared by incubation of negatively charged polystyrene microparticles with polyclonal rabbit IgGs, were determined by a variety of experimental techniques. These comprised dynamic light scattering (DLS), laser Doppler velocimetry (LDV) and atomic force microscopy (AFM). The particle diffusion coefficient, the hydrodynamic diameter, the electrophoretic mobility, the zeta potential and the suspension stability were determined as a function of pH for different ionic strengths.

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Immunolatex particles (LAmP-s), which were prepared by covalently co-immobilizing antiferritin and a mixture of pentaethylenehexamine-ended poly(ethylene glycol) (N6-PEG) of two different chain lengths onto the surface of polystyrene submicroparticles, were formulated with various antiferritin loads. Following the quantification of the bound antiferritin, as well as the differentiation of the physically adsorbed antiferritin from chemically bound antiferritin, using the copper reduction/bicinchoninic acid reaction (the Micro BCA method), dynamic light scattering (DLS) and electrophoretic mobility (mu(e)) measurements were performed to characterize the size, homogeneity, and surface charge of the complex. Compared to the control immunolatex particles, which were prepared similarly but traditionally using bovine serum albumin (BSA) as a blocking agent (LAB-s), the LAmP-s complex showed a difference only in the surface charge property, because of the altered surface treatment in the case of the LAmP-s (PEGylation) and LAB-s complexes (BSA covering).

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