Менингококковая поливакцина на основе очищенной IgAI протеазы

IgA1-protease allocated from the culture N. meningitidis serogroup A. As original materials were used three different intermediate products of vaccine production: cultural fluid, cetavlon supernatant and cetavlon precipitate. IgA1-protease was used to evaluate their protectivity and immunogenity. It was shown, that isolated IgA1 protease from the meningococcus serogroup A is able to protect mice, infected by meningococcus serogroup B.IgA1 протеаза, выделенная методами препаративной химии из промежуточных продуктов очистки менингококковой полисахаридной серогруппы А вакцины, обладает выраженной иммуногенной и протективной активностью для лабораторных мышей, обеспечивая их защиту от смертельного заражения вирулентными штаммами серогрупп А и В менингококков. Протективный эффект и нарастание анти-IgA1 антител позволяют прогнозировать вакцинные потенции изучаемого препарата, который может рассматриваться, в первую очередь, в качестве кандидата отсутствующей до настоящего времени менингококковой серогруппы В вакцины

Менингококковые вакцины: от капсульных полисахаридов до протеаз

Microbial capsular polysaccharides for many years provided a highly practical public health vaccines for preventing meningococcal, pneumococcal and Haemophilus influenza infection, and typhoid fever. Their application in the form of conjugates with protein carriers eliminate the gap in protection against these infections in children under one year. Extremely promising turned out offered us a new generation of vaccines, which have synthetic peptides conjugated to a meningococcal polysaccharide. Thus, new approaches to the solution of the problem of meningococcal disease vaccination serogroup B were open. In recent years, Russian researchers first suggested to use IgA1 protease (one of the major virulence factors of microbes and almost identical for mentioned below infections) for prevention of such diseases as meningococcal of all serogroups, pneumococcus and hemophilia infections. Patented processes for producing of the vaccine define domestic priority of its production and use.Микробные капсульные полисахариды на протяжении многих лет обеспечивали практическое здравоохранение высокоэффективными вакцинами для профилактики менингококковых, пневмококковых, гемофильных инфекций и брюшного тифа. А их применение в виде конъюгатов с белковыми носителями ликвидировало пробел при защите от этих инфекций детей в возрасте до одного года. Необычайно перспективным оказалось предложенное нами новое поколение вакцин, где уже синтетические пептиды конъюгировались с полисахаридами менингококков. Таким образом, были открыты подходы к решению проблемы вакцинопрофилактики менингококковой инфекции серогруппы В. В последние годы отечественные исследователи впервые предложили использовать для профилактики менингококковых инфекций всех серогрупп, а также инфекций, обусловленных пневмококками и гемофилами, один из главных факторов вирулентности этих микробов — IgA1 протеазу, практически идентичную для всей указанной группы инфекций. Запатентованные способы получения этой вакцины определяют отечественный приоритет ее производства и применения

Isolation and determination of the activity of IgA1 protease from Neisseria meningitidis

A method of the isolation and purification of IgA1 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 precipitate 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 developed. 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 μg. It was shown that IgA1 protease isolated from serogroup A meningococcus is capable of protecting experimental animals (mice) infected with meningococcus of serogroup B. © 2010 Pleiades Publishing, Ltd

Isolation and determination of the activity of IgA1 protease from Neisseria meningitidis

A method of the isolation and purification of IgA1 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 precipitate 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 developed. 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 μg. It was shown that IgA1 protease isolated from serogroup A meningococcus is capable of protecting experimental animals (mice) infected with meningococcus of serogroup B. © 2010 Pleiades Publishing, Ltd

Recombinant IgA1 protease from N. meningitidis. obtaining and properties

On the base of nucleotide sequence, coding IgA1 protease from Neisseria meningitidis serogroup B, strain ÌÑ58, which was determined from data base http://www.ncbi.nlm.nih.gov/Genbank), recombinant plasmide DNA was created comprising nucleotide sequence of IgA1 protease, strain H44/76, providing IgA1 protease expression in the host-cell (pBIGAPS1). The method of expression, isolation, purification and refolding of recombinant enzyme was developed. IgA1 protease exhibits high specificity and cleaves only IgA1, but not IgG. Immunogenic and protective properties of IgA1 protease to meningococcus of serogroup À, Â and Ñ were shown. Obtained enzyme can be considered as perspective polyvaccine candidate for prophylaxis against meningococcus infection, induced by bacteria N. meningitidis and especially against serogroup B

Recombinant IgA1 protease from N. meningitidis. obtaining and properties

On the base of nucleotide sequence, coding IgA1 protease from Neisseria meningitidis serogroup B, strain ÌÑ58, which was determined from data base http://www.ncbi.nlm.nih.gov/Genbank), recombinant plasmide DNA was created comprising nucleotide sequence of IgA1 protease, strain H44/76, providing IgA1 protease expression in the host-cell (pBIGAPS1). The method of expression, isolation, purification and refolding of recombinant enzyme was developed. IgA1 protease exhibits high specificity and cleaves only IgA1, but not IgG. Immunogenic and protective properties of IgA1 protease to meningococcus of serogroup À, Â and Ñ were shown. Obtained enzyme can be considered as perspective polyvaccine candidate for prophylaxis against meningococcus infection, induced by bacteria N. meningitidis and especially against serogroup B