Detection of Hemolytic Activity of Brucella melitensis Antigens Loaded in Nanocarriers

Abstract

The hemolytic potential of NPs has been the subject of numerous studies as a critical indicator of their safety in vaccine applications. In this study, the elemental compositions of Brucella melitensis Antigens (BMAs) loaded in Ca, Phosphate/ Chitosan nanocomposite created by encapsulation method. In addition, size, and surface topography were characterized by UV-VIS spectrophotometers, Energy Dispersive X-ray (EDS/EDX), and Atomic Force Microscopy (AFM) techniques. Furthermore, the cytotoxicity of Brucella melitensis antigens loaded in calcium phosphate/chitosan nanoparticles was determined by measuring the hemoglobin released from rabbit erythrocytes using a spectrophotometer at 545 nm. The results showed that λ max of BMAs, Ca, P/CsNPs were present at 1.987 at the wavelength 232 nm which differed from the absorbance values of CaP/Cs NPs and Brucella melitensis Antigens samples respectively. The main components of BMAs, Ca, P/CsNPs were C, O, and sodium, with 22.2%, 46.1%, and 20.7 % respectively and the secondary components were Ca and P with 4.4% and 1.3 % respectively. Additionally, other elements confirmed the presence of Brucella melitensis antigens in. nanocomposite. The average particle size was 30.15 nm and a total height of 107.4 nm. Additionally, the hemolysis index of BMAs, Ca, P/Cs NPs was not detectable against rabbit RBCs.  We can conclude that the unique properties of CaP/CsNPs enabled the carry of Brucella melitensis antigens and were nontoxic for RBCs, suggesting potential candidates for vaccine applications against Brucella melitensis.