Purification and performance evaluation of polyclonal antibody against the SARS-CoV-2
Purification and performance evaluation of polyclonal antibody against the SARS-CoV-2
Fatemeh Yaghoobizadeh,1,*Mohammad Roayaei Ardakani,2Mohammad Mehdi Ranjbar,3Mohammad Khosravi,4Hamid Galehdari,5
1. Department of Biology, Shahid Chamran University of Ahvaz, Ahvaz, Khouzestan, Iran. 2. Department of Biology, Shahid Chamran University of Ahvaz, Ahvaz, Khouzestan, Iran. 3. Razi Vaccine and Serum Research Institute, Karaj, Alborz, Iran. 4. Department of Pathobiology, Shahid Chamran University of Ahvaz, Ahvaz, Khouzestan, Iran. 5. Department of Biology, Shahid Chamran University of Ahvaz, Ahvaz, Khouzestan, Iran.
Introduction: SARS-CoV-2 is amongst the human coronaviruses which cause severe pneumonia or even death. Due to the importance of its rapid identification/treatment and with the regard to this fact that pAbs are relatively cheap, maybe able to neutralize antigens, this study was done with the purpose of production/purification of anti-SARS-CoV-2 polyclonal antibodies (pAb) in rat.
Methods: The purification of viral antigens was done by PEG/NaCl precipitation. Immunization was done by the purified antigen and ammonium sulfate precipitation (33%) was utilized for the purification of primary antibody. Following the final purification by ion-exchange chromatography, the the antibody’s specificity was verified by ELISA and western blotting (WB). All data were analyzed by SPSS software.
Results: The results showed that increase of PEG concentration, will increase the amount of concentrated virus. Besides, high-titres of pAb were induced in rat and a good antibody-antigen interaction was indicated in ELISA/WB results.
Conclusion: Herein, we presented a cheap and simple protocol for purification of relatively high quality of whole viral particles. The results verified that these antigens elicit a good immune response and have a good specificity against the CoV’s antigens.
Keywords: SARS-CoV-2; western blotting; polyethylene glycol; ELISA.