Production and Purification of Anti-Rhombomys opimus Immunoglobulins

Background: zoonotic cutaneous leishmaniasis (zcl) is an increasing public health problem in some endemic re- gions. horseradish peroxidase (hrp) conjugated rabbit anti-rhombomys opimus (r. opimus) ig is needed for im- munoblotting 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 in- jected into rabbit to produce anti-r. opimus ig antibody. the titration of antibody against r. opimus ig in rabbit se- rum was checked using indirect elisa. rabbit anti-r. opimus ig was purified by sepharose-4b-r. opimus ig column. re- activity 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.