IDENTIFICATION OF IMMUNOGENIC PROTEINS OF STRAINS OF BACILLUS ANTHRACIS IN MALDI TOF MS

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.

Bacillus anthracis, MALDI TOF MS, Bacillus anthracis, immunodominant proteins, dimensional electrophoresis, immunoblotting, MALDI TOF MS