Production and purification of anti-rhombomys opimus immunoglobulins
Iranian Journal of Arthropod-Borne Diseases. 2011; 5 (2): 69-76
in English
| IMEMR
| ID: emr-132747
ABSTRACT
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. 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. 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 18000 using direct ELISA. 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
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Index:
IMEMR (Eastern Mediterranean)
Language:
English
Journal:
Iran. J. Arthropod-Borne Dis.
Year:
2011
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