CpG-Containing Oligodeoxynucleotides and Freund Adjuvant in Combination with  Alum Augment the Production of Monoclonal Antibodies Against Recombinant HBsAg

Mahsa i  Khayyati Kohnehshahr; Nowruz  Delirezh; Leili Aghebati Maleki

doi:10.18502/ajmb.v14i2.8880

Mahsa i Khayyati Kohnehshahr Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran
Nowruz Delirezh Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran
Leili Aghebati Maleki Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

DOI: https://doi.org/10.18502/ajmb.v14i2.8880

Keywords: Aluminum sulfate, Freund adjuvant, Hepatitis B surface antigens, Monoclonal antibodies

Abstract

Background: Adjuvants are essential to potentiate the immune response to inoculated antigens and play a central role in vaccine development. Alum is generally used as a classic adjuvant, although it does not stimulate proper immunity, and some of the immunized subjects have low or no antibody response. Efforts have been continued to find more efficient adjuvants for better antibody responses. In the present study, the efficacy of three formulations of adjuvants, i.e. Cysteine p Guanine Oligodeoxynucleotide (CpG ODN), alum, and Freund, in the production of monoclonal anti Hepatitis B Surface Antigen (HBsAg) antibodies was investigated.

Methods: To immunize mice, regular hepatitis B vaccine containing recombinant HBsAg and alum was used with CpG ODN or Freund adjuvants, and splenocytes of hyperimmunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by Enzyme-Linked Immunosorbent Assay (ELISA) using HBsAg as coating antigen followed by a limited dilution process.

Results: The results showed that by using all three formulations of adjuvants, monoclonal antibody (mAb) specific to HBsAg was successfully generated. It was also found that the mice immunized with (HBsAg + Alum) + CpG had the highest concentration of antibody production in serum and hybridoma supernatants as well as positive clones. Based on these findings, the addition of CpG ODN also induced a higher antibody response compared with Complete Freund’s Adjuvant (CFA).

Conclusion: Results of this study showed that CpG and Freund adjuvants could be efficient partners for alum in the immunization period of the process of monoclonal antibody production.