Development of standardization approaches for recombinant endolysin LYSECD7-based pharmaceutical substance

Relevance. Lytic bacteriophages endolysins represent one of the most promising classes of antibacterial agents in the post-antibiotic era. Since this class of antibacterials is quite new there is an urgent need for development of approaches to the endolysins pharmaceutical substances standardization. Objective of the study was to develop the approaches to the recombinant endolysin LysECD7 pharmaceutical substance standardization. Materials and methods. The identity and purity of the endolysin pharmaceutical substance was assessed by SDS-PAGE in 16% polyacrylamide gel. The specific antibacterial activity of the LysECD7 pharmaceutical substance was determined using a novel microbiological method counting the CFU reduction of the Acinetobacter baumannii Ts 50-16 test strain after the endolysin exposure. The sterility of the LysECD7 pharmaceutical substance was determined by the membrane filtration method. The bacterial endotoxins level was determined by a chromogenic endpoint test. The quantity of the protein in the pharmaceutical substance was determined by the colorimetric method with bicinchoninic acid. Results. During the work the approaches to the standardization of the endolysin LysECD7 pharmaceutical substance were developed including the basic quality attributes such as "Electrophoresis in polyacrylamide gel" and "Specific activity" to confirm the identity, "Sterility", "Bacterial endotoxins" and "Quantification". Conclusions. Approaches to the standardization of the LysECD7 endolysin pharmaceutical substance were developed including such quality attributes as "Identity", "Purity" and "Quantification", which allow to reach the stable efficiency and safety of the pharmaceutical substance aiming to develop innovative effective antibacterial drugs.

LysECD7, Endolysins, LysECD7, Gram-negative bacteria, pharmaceutical substance, standardization, quality control