Preparation and preclinical evaluation of two novel Staphylococcus aureus capsular polysaccharide 5 and 8-fusion protein (Hla-MntC-SACOL0723) immunoconjugates

Abstract

Staphylococcus aureus is one of the most common pathogens in the hospital and the community. The emergence of broad-spectrum antibiotic resistance in S. aureus has made the treatment process more difficult. Therefore, it is obvious that an effective prevention strategy against the pathogen could significantly reduce costs related to care in hospitals. In this report, we describe a simple approach to conjugate S. aureus capsular polysaccharide 5 (CP5) from S. aureus Reynolds strain and 8 (CP8) from S. aureus Becker strain to a fusion protein (Hla-MntC-SACOL0723) and investigation of its bioactivity. The conjugation was done by using ADH (as a bridge) and EDAC (as a coupling agent). The immunoconjugates were characterized by routine polysaccharide/protein contents assays followed by reverse phase chromatography and FTIR spectroscopy. The groups of mice were immunized with conjugate vaccines, capsular polysaccharides, and phosphate-buffered saline (PBS) as a control group. The functional activity of the vaccine candidates was evaluated by ELISA, opsonophagocytosis tests, and determination of bacterial load in challenge study. The results showed that the specific antibody (total IgG) titers raised against conjugate molecules were higher than those of the nonconjugated capsular polysaccharides. The opsonic activity of the conjugate vaccines antisera was significantly higher than polysaccharides alone (58% reduction in the number of bacteria versus 16.3% at 1:2 dilution, p