ABSTRACT
Gold nanoparticle conjugates with Vibrio cholerae antigens were synthesized. The animals were immunized with the obtained conjugates. Rabbit polyclonal antibodies to the antigens were obtained, which showed high specific activity. On the model of white laboratory mice, the protective activity of conjugates of cholera antigens with nanoparticles during infection of vaccinated animals was evaluated using a commercial vaccine as a control. It was shown that in terms of immunogenicity, the created prototypes of cholera vaccine using gold nanoparticles as a carrier and adjuvant complied with the production regulations for the Russian national cholera chemical vaccine.
Subject(s)
Antibodies, Bacterial/immunology , Antigens, Bacterial/immunology , Cholera Vaccines/immunology , Cholera/immunology , Cholera/prevention & control , Gold/chemistry , Metal Nanoparticles/chemistry , Animals , Antigen-Presenting Cells , Antioxidants/chemistry , Chinchilla , Immunization , Mice , Nanotechnology/methods , Reproducibility of Results , Vaccines, Attenuated/immunology , Vibrio cholerae/immunologyABSTRACT
The aim of this research was to design a method of immobilization of high-purity human butyrylcholinesterase on the surface of gold nanoparticles preserving the activity of the enzyme. In order to achieve this aim, the method of fractionation and purification of human butyrylcholinesterase from plasma was modified. The synthesis of 15-nm gold nanoparticles was carried out by citrated method. A method of conjugation of the high-purity butyrylcholinesterase with gold nanoparticles was developed. It was found that the Immobilization of butyrylcholinesterase on the surface of gold nanoparticles resulted in a significant (to 23%) increase in the specific activity of the enzyme.
Subject(s)
Butyrylcholinesterase , Gold Compounds/chemical synthesis , Metal Nanoparticles , Butyrylcholinesterase/chemistry , Butyrylcholinesterase/isolation & purification , Enzyme Stability , Gold Compounds/chemistry , Humans , Hydrogen-Ion Concentration , Metal Nanoparticles/chemistry , Particle SizeABSTRACT
The structural influence of Azospirillum lipopolysaccharides (LPS) and lipopolysaccharide-protein complexes (LPPC) on carrot, erythrocyte, and bacterial cell suspensions was explored. The structural potentialities of O-specific polysaccharide fragments of LPS and protein fractions of LPPC were also evaluated. An ability to induce the formation of three kinds of structures in the cell suspensions was revealed depending on the chemical composition of the preparations used. The first and the second ones were connected with effects of cell aggregation (a relatively fast process) and agglutination (a relatively slow process). The third one resulted in phase separation of erythrocyte suspensions (a medium-speed process), with segregating the cells to a separate homogeneous liquid phase.
Subject(s)
Azospirillum brasilense/metabolism , Bacterial Proteins/chemistry , Lipopolysaccharides/chemistry , Agglutination , Azospirillum brasilense/physiology , Bacterial Capsules/chemistry , Cell Aggregation/physiology , Daucus carota/cytology , Erythrocytes/cytology , Symbiosis/physiologyABSTRACT
We have devised a protocol for the isolation and identification of a proliferative antigen of the initial cells of wheat stem meristems (termed PAI). We have carried out a variety of immunochemical and immunocytochemical methods, using colloidal gold (CG) complexed with monospecific antibodies to PAI as the marker for the detection of PAI. We have been able to determine the effectiveness of immunoaffinity chromatography in isolating PAI from plant tissues and have shown the advantages of CG over enzyme labels for identification of the antigen. Finally, we have obtained a purified preparation of PAI and have determined its molecular mass (approximately 83 kDa).
Subject(s)
Antigens/analysis , Immunoblotting/methods , Meristem/immunology , Plant Stems/immunology , Triticum/immunology , Cell Division , Gold , Immunoenzyme Techniques/methodsABSTRACT
The obtainment of stable solutions of inverse problems for studying the disperse composition of suspensions using effects of elastic light scattering was discussed. Versions of a regularization of solving the inverse problems of the spectroturbidimetric method were considered, taking into account unavoidable restrictions on the scope of the necessary prior data for particles and on the width of the spectral interval for real biological disperse systems. Possibilities for increasing the number of particle parameters determined in a single optical experiment were analyzed. They were shown to be provided by the use of effects of the orientation ordering of a system on combination of capabilities of the methods of spectroturbidimetry and electro-optics, using bacterial cell suspensions as an example. © 1999 Society of Photo-Optical Instrumentation Engineers.