Tehran University of Medical Sciences Journal of Arthropod-Borne Diseases 2322-1984 5 2 2011 12 15 69 76 EN AA Akhavan Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran R Ghods Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran M Jeddi-Tehrani Department of Hybridoma, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran MR Yaghoobi-Ershadi Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran A Khamesipour Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran AR Mahmoudi Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran 2015 10 03 Background: Zoonotic cutaneous leishmaniasis (ZCL) is an increasing public health problem in some endemic re­gions. Horseradish peroxidase (HRP) conjugated rabbit anti-Rhombomys opimus (R. opimus) Ig is needed for im­munoblotting and ELISA tests used to explore the immune response of the rodents against the sand fly saliva. In this study, the production of HRP conjugated rabbit anti-R. opimus Ig was conducted for the first time.Methods: Rhombomys opimus Ig was purified from serum by protein G affinity chromatography column and in­jected into rabbit to produce anti-R. opimus Ig antibody. The titration of antibody against R. opimus Ig in rabbit se­rum was checked using indirect ELISA. Rabbit anti-R. opimus Ig was purified by Sepharose-4B-R. opimus Ig column. Re­activity of this antibody was assessed by indirect ELISA and was conjugated to HRP by periodate method. Results: Approximately 3.5 mg Ig was purified from 1 ml R. opimus serum using protein G affinity chromatography col­umn. The molecular weight of purified R. opimus Ig was estimated about 150 kDa by SDS-PAGE. Nearly 2.3 mg rab­bit anti-R. opimus Ig was purified from 1 ml immunized rabbit serum. The purified antibody was conjugated to HRP and the optimum titer of HRP conjugated rabbit anti-R. opimus Ig was determined as 1:8000 using direct ELISA. Conclusion: HRP conjugated rabbit anti-Gerbil IgG has been produced by a few companies, but to our knowledge HRP conjugated rabbit anti-R. opimus Ig is not commercially available. Production of HRP conjugated rabbit anti-R. opimus Ig is considerably helpful for immunological studies of R. opimus, the main reservoir host of ZCL in Iran as well as some other countries. https://jad.tums.ac.ir/index.php/jad/article/view/93 https://jad.tums.ac.ir/index.php/jad/article/download/93/91