production and purification of anti-rhombomys opimus immunoglobulins

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.