ИДЕНТИФИКАЦИЯ ИММУНОГЕННЫХ БЕЛКОВ ШТАММОВ BACILLUS ANTHRACIS В MALDITOF MS

M. P. Chervakova, T. Sharov, L. P. Barkova, A. Barkov, D. Viktorov, A. Toporkov
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引用次数: 1

Abstract

Aim. Identification of obtained in host-simulated conditions immunogenic proteins of isogenic variants of Bacillus anthracis 575/122. Materials and methods. We used culture filtrate of isogenic variants of B. anthracis 575/122: R02 (pXOL pXO2 + ); R01 (pXO1 + pXO2‘); R00 (pXOL pX02~), obtained in host-simulated conditions. In the one-dimensional polyacrylamide gel electrophoresis and immunoblotting with hyperimmune serums immunodominant proteins, that have been identified in MALDI TOF MS. Results. Immunoblotting revealed proteins with molecular masses in range 97 - 14.1 kDa. 90 kDa protein from strain B. anthracis 575/122 R01 in MALDI TOF MS was identified as protective antigen with 85.810 kDa. Protein with molecular mass 60 kDa was identified as GMP synthase with molecular mass 57.239 kDa. In the culture filtrates of three strains two common antigen were identified: protein with molecular mass 97 kDa, identified as B. anthracis EA 1 with molecular mass 91.361 kDa protein and 45 kDa protein as enolase B. anthracis with molecular mass 46.418 kDa. Conclusion. Thus, the conditions that simulate the host can promote the production of immunodominant proteins of B. anthracis. The data about molecular-weight characteristics of protective antigen and EA 1 protein as well as some of proteases of B. anthracis are confirmed by the MALDI TOF MS. The results can be used for isolation of these proteins to improve the diagnostic and vaccine preparations.
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的目标。在宿主模拟条件下获得的炭疽芽孢杆菌575/122等基因变异免疫原性蛋白的鉴定。材料和方法。采用炭疽芽孢杆菌575/122:R02 (pXOL pXO2 +)等基因变异培养滤液;R01 (px1 + pXO2 ');R00 (pXOL pX02~),在主机模拟条件下得到。在一维聚丙烯酰胺凝胶电泳和免疫印迹与高免疫血清免疫优势蛋白,已鉴定出MALDI TOF ms结果。免疫印迹显示蛋白分子量在97 - 14.1 kDa之间。MALDI TOF质谱鉴定鉴定出来自炭疽芽孢杆菌575/122 R01的90 kDa蛋白为保护抗原,分子量为85.810 kDa。分子量为60 kDa的蛋白被鉴定为分子量为57.239 kDa的GMP合成酶。在三株菌株的培养滤液中鉴定出两种共同抗原:分子量为97 kDa的蛋白,鉴定为炭疽芽胞杆菌EA 1蛋白,分子量为91.361 kDa, 45 kDa蛋白为炭疽芽胞杆菌烯醇化酶蛋白,分子量为46.418 kDa。结论。因此,模拟宿主的条件可以促进炭疽芽胞杆菌免疫优势蛋白的产生。MALDI TOF质谱法证实了炭疽芽孢杆菌的保护性抗原、EA - 1蛋白及部分蛋白酶的分子量特征,结果可用于分离这些蛋白,以提高诊断和疫苗制备水平。
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来源期刊
Zhurnal mikrobiologii, epidemiologii, i immunobiologii
Zhurnal mikrobiologii, epidemiologii, i immunobiologii Immunology and Microbiology-Immunology and Microbiology (miscellaneous)
CiteScore
1.40
自引率
0.00%
发文量
51
审稿时长
8 weeks
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