Razi Vaccine & Serum Research Institute Co-Publisher: Negah Scientific Publisher Archives of Razi Institute 0365-3439 78 4 2023 08 01 Effects of Adjuvant and Immunization Route on Antibody Responses against Naja Naja oxiana Venom تاثیر ادجوانت و روش ایمن سازی برروی میزان آنتی بادی ایجاد شده برعلیه زهر مار ناجا ناجا اکسیانا 1177 1183 128775 10.32592/ARI.2023.78.4.1177 EN Sh Kashkooli Department of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran S Khamehchian Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran M Dabaghian Department of Immunology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran M Namvarpour Department of Immunology, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran M Tebianian Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran Journal Article 2023 01 21 <em>Naja naja oxiana</em> (NNO) is one of the important venomous species in Iran. The current snakebite treatment is antivenom therapy that deals with hyper immunization of horses with crude or fractionated snake venom plus traditional adjuvants, like Freund’s adjuvant. For improvement of antivenom production, it has been suggested to use different adjuvant systems or immunization procedures. In this study, humoral immune responses against immunogenic fractions of NNO venom (NNO3 and NNO4) and crude venom have been compared by usage of different adjuvant and immunization routes. Additionally, a new indirect enzyme-linked immunosorbent assay (ELISA) was set up for the detection of specific antivenom antibodies. This study was conducted on six different groups of female Dutch rabbits that were hyperimmunized using crude and fractionated NNO venom, along with Freund’s and MF59 adjuvants through subcutaneous or intramuscular route. The immunization was performed four times with 10-day intervals and the levels of specific antibodiees were detected by indirect ELISA. The statistical analysis reveals a negligible variation in the antivenom titers among the venom-inoculated groups, regardless of the adjuvant type or the immunization route. Finally, it was concluded that the fractions are efficient for antivenom production, and it is possible to use MF59 adjuvant via subcutaneous routes as an alternative to Freund’s adjuvants considering its fair immunopotentiation capacity and safety in animals. https://archrazi.areeo.ac.ir/article_128775_21fd28cb0d67fe6066065acc97e5e8f0.pdf