Obtaining of native microbial antigens with anti-adhesive properties

Abstract

Annotation. Traditional methods of sanitation of bacterial carriers of nosocomial infections of staphylococcal genesis using antibacterial drugs are ineffective and lead to the formation of antibiotic resistance in strains of microorganisms. Currently, when developing new classes of immunobiological drugs, it is recommended to use pathogen-associated molecular structures (PAMS) of bacteria that block surface epitopes and trigger the mechanisms of innate and adaptive immunity. The aim of the work was to obtain native surface antigens of Staphylococcus aureus using physical factors. Quantification of protein content in the obtained antigen preparations was performed using spectrophotometry according to the method of O. H. Lowry et al. (1951). The content of teichoic acids was determined according to the method of A. R. Archibald (2012). Adhesive properties were studied according to the method of V. I. Brilis et al. (1986). Each sample was tested in three to four replicates. Data were statistically processed using Statistica 8.0 (StatSoft Inc., USA) or Excel 2003, 2010 (Microsoft, USA). S. aureus surface antigens (Ag) were obtained as follows: a test culture of S. aureus ATСС 25923 was pre-irradiated with 61.0 GHz millimeter frequency range waves for eight hours, and then sonicated with a GZ-109 (60 kHz) or UZIN-2T (44 kHz) instrument and the protein and teichoic acid contents were measured. It was found that pretreatment of suspension S. aureus ATСС 25923 by 61.0 GHz millimeter frequency range waves increased the protein content in the samples by 1.5-1.9 times (p<0.01). Lipoteichoic acid content exceeded ribiteichoic acid content, on average, by 16.7 times (p<0.001) in samples obtained with the UZIN device and by 28.5 times (p<0.001) in samples obtained with the G3-10 device. All of the antigenic (Ag) samples obtained had anti-adhesive activity against S. aureus ATСС 25923, with the Ag5 sample (obtained using the GZ-109 instrument) exhibiting a more pronounced anti-adhesive activity. The use of physical factors will contribute to the standardization of the process of obtaining surface native structures and reduce the toxicity and reactogenicity of immunobiological preparations.