The method for obtaining of the protective pertussis antigen by low-frequency ultrasound
Keywords:ultrasound, fraction, antigen, vaccine, immunogenicity, Bordetella pertussis
This article describes the physical technology for obtaining surface antigens with protective properties from Bordetella pertussis microbial cells in the absence of additional use of chemical and synthetic substances. The mechanical destruction of cell membranes of microorganisms was carried out under mild conditions using low power low-frequency ultrasound, it allowed not to damage the protective bioactive substances. Ultracentrifugation of microbial ultrasound desintegrates and the subsequent gel – chromatographic separation allowed to obtain a protective antigen with the molecular weight of ~ 8.1 kDa and significantly enhance its specific weight from (62.5±9.1%) to (86.2±4.6%) (P<0.05). The study of toxicity in the test of the mice weight changes showed the presence of pathogenicity factors in the entire antigenic complex and in the fractionated component with the weight of ~ 3.0 kDa. And the purified native antigen with the weight of ~ 8.1 kDa in the calculated dose (160 mg) did not contain toxic compounds. The antigen with the molecular weight of ~ 8.1 kDa showed no histamine-sensitizing and dermonecrotic properties. While studying immunogenicity of the fraction with the weight of ~ 8.1 kDa a strong direct correlation was found between increase of vaccination antigen doses and the corresponding increase of the immunity intensity of vaccinated animals. It indicates the specificity of the results obtained and the protective activity of this fraction. The percentage of survived mice vaccinated by the native antigen with the molecular weight of ~ 8.1 kDa 1.9 times exceeded the percentage of animals vaccinated by the standard industry sample of pertussis vaccine.
Аналітична нормативна документація на кашлюкову суспензію: [методичні вказівки] / наказ № 23. – 24.07.2003. – ЗАТ «Біолік», 2003. – 32 с.
Гельман В. Я. Медицинская информатика: [практикум] / В. Я. Гельман. – СПб: Питер, 2002. – 480 с. – (Серия «Национальная медицинская библиотека»)
Гланц С. Медико–биологическая статистика / С. Гланц : [пер. с англ.]. – М. : Практика, 1998. – 459 с.
Методическое руководство по лабораторной оценке качества бактерийных и вирусных препаратов (вакцины, анатоксины, сыворотки, бактериофаги и аллергены) : [методическое руководство] / отв. редактор С. Г. Дзагуров : Москва, 1972. – 301с.
Сова В.В. Выделение и очистка белков. Методическое пособие по курсу "Химия и биохимия белков и ферментов": [методическое пособие] / В.В. Сова, М.И. Кусайкин. - Владивосток: узд-во Дальневост. Ун-та, 2006.- 42 с.
Lowry O. H., Rosebrough O. H., Farr N. J. [et al.] / J Biol Chem. – 1951.– № 193.– Р. 265–275.
WHO Working Group meeting on Standardization of Acellular Pertussis Vaccines: potency assay. – 7–9 November 2007. [Еlectronic resource] – Access mode: http://www.who.int/ru.
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