Salmonella typhimurium is the primary causative agent of salmonellosis in animals and humans. Many methods are used to treat salmonellosis including our previous study in which orally administrated camel milk to infected mice reduced their deaths and reduce bacterial load in their blood, livers and spleens.

Objectives: To investigate the ability of purified IgG₃ antibodies from immunized camel to detect Salmonella typhimurium. And to study the invitro properties of these antibodies that may result in the protection against Salmonella infection.

Materials and methods: Two female camels were used; one as a control and the other was vaccinated subcutaneously with S. typhimurium. Camel IgG₃ antibodies were purified from collected sera and milk sampels. Then SDS-PAGE and immunoblotting used for antibody detection. The purified IgG₃ antibodies used for production of mouse anti-camel IgG₃ antibodies. Stability of IgG₃ antibodies was measured under different variables of temperature and pH. Purification and charecterization of urine components that are reactive with S. typhimurium also performed.

Results and discussion: Purified camel IgG₃ antibodies showed reactivity to ward S. typhimurium, even after conjugation with HRP. After hybridoma production; two different monoclonal antibodies isotypes were produced which are IgG₁ and IgG₃. These antibodies had high specificity for camel IgG₃ as revealed by immunoblotting of camel sera and milk. Furthermore, camel IgG₃ antibodies showed high resistance to extreme pH values as they retained 90% of their binding activity after 2 h incubation at pH 2. They also showed significant resistance to thermal denaturation at high temperatures (>70^○C). Interestingly, these IgG₃ antibodies conserved 62 % of their binding activity after one week incubation at room temperature and 55% after 21 days. Finally, our results proved the presence of antibodies that showed reactivity toward S. typhimurium in camel urine.

Conclusion: The heavy chain antibodies particularly IgG3 in this study found to resist thermal, pH and pepsin denaturation. Which in turn enable them to be suitable normally occurring molecules for passive immunization against S. typhimurium especially via oral route.

Conference Title

Palestinian International Conference in Medical Lab Sciences (Ramallah, August 14-16, 2014)

Conference Country

Palestine

Conference Date

Dec. 14, 2013 - Dec. 14, 2013

Conference Sponsor

نقابة الطب المخبري الفلسطينية