Production and Purification of CAMP-Sialidase Chimer Protein as a Vaccine Candidate for Acne and Its Immunization in the Animal Model

Mansouri, Kowsar; Ahmadi, Ali; Salimian, Jafar; Zargham, Parvin; Amani, Jafar

doi:10.30491/jabr.2020.243402.1270

Production and Purification of CAMP-Sialidase Chimer Protein as a Vaccine Candidate for Acne and Its Immunization in the Animal Model

Document Type : Original Article

Authors

Kowsar Mansouri ¹

Ali Ahmadi ²

Jafar Salimian ³

Parvin Zargham ⁴

Jafar Amani ⁵

¹ New Hearing Technologies Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

² Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

³ Chemical Injuries Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

⁴ Department of Molecular and Cellular Sciences, Faculty of Advanced Sciences and Technology, Pharmaceutical Sciences Branch, Islamic Azad University (IAUPS), Tehran, Iran

⁵ Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran

10.30491/jabr.2020.243402.1270

Abstract

Introduction: Acne vulgaris affects ~85% of young adults aged 12–25 years. Regarding the critical role of Propionibacterium acnes in the pathogenesis of acne and current therapeutic failures, developing efficient acne vaccines are appreciated. The purpose of this study was to design a chimeric protein from CAMP and sialidase parts of Propionibacterium acnes and evaluating its immunogenicity in a mice model as an acne vaccine candidate.
Materials and Methods: CAMP-Sialidase recombinant gene expression was carried out through cloning in the vector pET28a and transferring to E.coli BL21DE3. The protein was purified using the Ni-NTA column and its concentration was measured. The recombinant protein was injected in test and control mice groups. Then, antibody titration and challenge tests were made to determine the immunogenicity.
Results: After successful expression and purification, the protein band was observed at a molecular weight of 65 kDa. Western blotting confirmed the purified protein. The results of serum ELISA indicated the IgG titer was 1:204800 and 1:1600 against the recombinant protein and inactivated P. acne, respectively. Although there was no change in test mice, inflammation happened in 50% of the control group.
Conclusions: The current study demonstrated that the CAMP-Sialidase recombinant protein can appropriately induce humoral antibodies. However, more evaluations need to introduce it as an acne vaccine candidate.

Keywords

Acne

Propionibacterium acnes

CAMP-Sialidase Recombinant Protein

Vaccine