鼠疫亚单位和立克次体疫苗制剂中蛋白质抗原的直接定量

Q3 Medicine
P. Kopylov, S. V. Dentovskaya
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引用次数: 0

摘要

本工作的目的是提出直接定量测定制剂和各种亚单位疫苗原型中鼠疫耶尔森菌和拉氏立克次体蛋白抗原含量的方法。材料和方法。将鼠疫杆菌LcrV和Caf1抗原分别包埋在分子微胶囊化鼠疫疫苗(MMPV)的物质中和与鼠疫蛋白共沉淀的氨基酸微晶中作为模型抗原。将R.raoultii Adr2、OmpB24和YbgF抗原吸附在立克次体疫苗的原型物质上。从MMPV微胶囊中释放鼠疫抗原是通过分别用有机溶剂、二氯甲烷和甲醇连续处理后者来进行的;将载体微晶溶解在pH 6.0的0.1M柠檬酸钠缓冲液中。立克次体疫苗的原型物质中的抗原含量是通过测量未与铝凝胶结合的蛋白质的量来确定的。通过处理聚丙烯酰胺凝胶的数字图像来计算表征疫苗制剂的物质和原型中抗原含量的定量参数,聚丙烯酰胺凝胶是通过从载体中提取的蛋白质-抗原组分的电泳获得的。结果和讨论。研究了从基于不会引起蛋白质降解的氨基酸微晶和聚乳酸微胶囊的亚单位疫苗制剂中直接提取和随后定量分析鼠疫杆菌LcrV和Caf1抗原的方法。已经确定了LcrV和Caf1在微晶物质中结合的不同性质,而从微晶释放的抗原的比例只有在它们完全溶解的情况下才被量化。研究发现,在从微晶中提取的低浓度LcrV和Caf1蛋白下,有必要浓缩提取物并随后去除盐,以使其可靠地可视化。已经证实,在200μl悬浮液的剂量体积中,10μg拉乌尔蒂猪瘟抗原和蛋白质足以用于电泳法的定量分析。讨论了替代直接提取抗原的其他物理化学方法用于评估疫苗制剂的组成和质量的前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Direct Quantification of Protein Antigens in Subunit Plague and Rickettsial Vaccine Preparations
The aim of the work was to put forward the methods for direct quantitative determination of the content of Yersinia pestis and Rickettsia raoultii protein antigens in preparations and various prototypes of subunit vaccines. Materials and methods. Y. pestis LcrV and Caf1 antigens enclosed in the substance of the molecular microencapsulated plague vaccine (MMPV) and separately, in microcrystals of amino acids co-precipitated with plague proteins were used as model antigens. R. raoultii Adr2, OmpB24, and YbgF antigens were adsorbed on the prototype substance of the rickettsia vaccine. The release of plague antigens from MMPV microcapsules was carried out through successive treatment of the latter with organic solvents, methylene chloride and methanol, respectively; the carrier microcrystals were dissolved in 0.1 M sodium citrate buffer at pH 6.0. The antigen content in the prototype substance of the rickettsial vaccine was determined by measuring the amount of proteins not bound to the alumogel. Quantitative parameters characterizing the content of antigens in the substances and prototypes of vaccine preparations were calculated by processing digital images of polyacrylamide gels obtained by electrophoresis of protein antigen fractions extracted from carriers. Results and discussion. Methods for direct extraction and subsequent quantitative analysis of Y. pestis LcrV and Caf1 antigens from subunit vaccine preparations based on amino acid microcrystals and polylactide microcapsules that do not cause protein degradation have been studied. A different nature of the binding of LcrV and Caf1 in the substances of microcrystals has been established, while the proportion of antigens released from microcrystals has been quantified only in case of their complete dissolution. It was found that at low concentrations of LcrV and Caf1 proteins extracted from microcrystals, it is necessary to concentrate the extracts with subsequent removal of salts for their reliable visualization. It has been confirmed that 10 μg of plague antigens and proteins of R. raoultii in a dose volume of 200 μl of suspension is sufficient for quantitative analysis using electrophoretic method. The prospects of other physicochemical methods alternative to direct extraction of antigens for evaluating the composition and quality of vaccine preparations are discussed.
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来源期刊
Problemy Osobo Opasnykh Infektsii
Problemy Osobo Opasnykh Infektsii Medicine-Infectious Diseases
CiteScore
1.90
自引率
0.00%
发文量
79
审稿时长
12 weeks
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