RESUMEN
The study of enzymatic and protective properties of recombinant IgA1 protease in active and mutant form showed that active form of IgA1 protease exhibited species - and type-specificity for mouse and human immunoglobulins. Mutant form, which did not exhibit enzymatic activity, had protective properties against meningococcal infection, induced by meningococcus serogroup A, B and C protecting the mice from lethal infection by living virulent culture of heterologous serogroups of meningococcus. Obtained results make it possible to consider IgA1 protease as a perspective preparation at the stages of development of polyvalent vaccine for protection the people from meningococcal infection of various etiology.
Asunto(s)
Proteínas Bacterianas/inmunología , Infecciones Meningocócicas/prevención & control , Vacunas Meningococicas/inmunología , Neisseria meningitidis/inmunología , Serina Endopeptidasas/inmunología , Animales , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/genética , Protección Cruzada , Femenino , Humanos , Inmunización , Infecciones Meningocócicas/inmunología , Vacunas Meningococicas/administración & dosificación , Ratones , Ratones Endogámicos BALB C , Mutación , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Serina Endopeptidasas/administración & dosificación , Serina Endopeptidasas/genética , Serotipificación , Vacunas de SubunidadRESUMEN
A method of the isolation and purification of IgAl protease from a culture of Neisseria meningitidis serogroup A has been developed. Three inactivated intermediates of the production of the meningococcal vaccine, a culture liquid, as well as a supernatant and sediment obtained by the precipitation of bacterial cells by cetavlon, served as a starting material. The purity of IgA1 protease was determined by SDS-PAGE. An immunoenzyme assay for determining the IgA1 protease activity has been devised. The yield of the enzyme with a specific activity of 0.5 to 4 million units/mg from 103 g of the cetavlon precipitate (40 l of culture liquid) was about 600 mug. It was shown that IgAl protease isolated from serogroup A meningococcus is capable of protecting experimental animals (mice) infected with meningococcus of serogroup B.
Asunto(s)
Inmunoglobulina A/metabolismo , Neisseria meningitidis/enzimología , Serina Endopeptidasas/metabolismo , Animales , Meningitis Meningocócica/inmunología , Meningitis Meningocócica/prevención & control , Vacunas Meningococicas/inmunología , Ratones , Ratones Endogámicos BALB C , Neisseria meningitidis/fisiología , Serina Endopeptidasas/inmunología , Serina Endopeptidasas/aislamiento & purificaciónRESUMEN
A composite sorbent based on porous glass beads modified with thin polyaniline coating was prepared by precipitating aniline polymerization in the presence of carrier particles. It was shown that the modification ensures the uniform coating of the inner surface of the carrier pores with the polymer layer approximately 70 A thick. It was shown that the resulting material retains the initial porosity of the carrier and is selective in the separation of nucleic acids and proteins. The polyaniline-coated sorbents were shown to be efficient for both the preparative DNA isolation from bacterial lysates and for analytical purposes, in particular, for studying DNA fragmentation during apoptosis proceeding under UV irradiation of cell lysates of colon carcinoma. The morphological and chromatographic characteristics of the new sorbent were demonstrated to be similar to those of the polyfluorobutadiene sorbent.