Mojtaba Sankian1, Yaser Bagheri1, Fatemeh Vahedi2, Farahzad Jabbari Azad3, Abdol-Reza Varasteh1. 1. Immunobiochemistry Lab, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran. 2. Razi Vaccine & Serum Research Institute, Mashhad, Iran. 3. Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Abstract
BACKGROUND: Allergy is a clinical disorder affecting humans worldwide. Allergenic extracts prepared from natural source materials remain heterogeneous in composition and content, but are regularly used for diagnosis and immunotherapy. Recombinant allergens are suitable candidates to use in place of natural allergens; however, the recombinant allergens should be assessed and compared with the natural ones. Cuc m 2 (profilin), one of the most important allergens of melon (Cucumis melo), has been cloned and was expressed in Escherichia coli (E. coli). We aimed to evaluate the validity of recombinant Cuc m 2 (rCuc m 2) in the diagnosis of melon allergy and investigate whether rCuc m 2 could be used as a replacement for natural Cuc m 2 (nCuc m 2). METHODS: nCuc m 2 was purified by immuno-affinity chromatography and rCuc m 2 was purified by metal-affinity chromatography. SDS-PAGE and western blotting were carried out to evaluate the purification methods. Skin prick tests (SPT), and enzyme immunoassays to determine specific IgE, were performed with the natural and recombinant purified allergens on 53 patients with melon allergy. RESULTS: rCuc m 2 elicited no significantly different responses in skin compared with nCuc m 2. All patients' sera showed similar ODs in ELISAs with natural and recombinant profilin. CONCLUSION: rCuc m 2 evoked strong immuno-reactivity equivalent to nCuc m 2, and has potential for diagnosis of melon allergy.
BACKGROUND:Allergy is a clinical disorder affecting humans worldwide. Allergenic extracts prepared from natural source materials remain heterogeneous in composition and content, but are regularly used for diagnosis and immunotherapy. Recombinant allergens are suitable candidates to use in place of natural allergens; however, the recombinant allergens should be assessed and compared with the natural ones. Cuc m 2 (profilin), one of the most important allergens of melon (Cucumis melo), has been cloned and was expressed in Escherichia coli (E. coli). We aimed to evaluate the validity of recombinant Cuc m 2 (rCuc m 2) in the diagnosis of melon allergy and investigate whether rCuc m 2 could be used as a replacement for natural Cuc m 2 (nCuc m 2). METHODS: nCuc m 2 was purified by immuno-affinity chromatography and rCuc m 2 was purified by metal-affinity chromatography. SDS-PAGE and western blotting were carried out to evaluate the purification methods. Skin prick tests (SPT), and enzyme immunoassays to determine specific IgE, were performed with the natural and recombinant purified allergens on 53 patients with melon allergy. RESULTS: rCuc m 2 elicited no significantly different responses in skin compared with nCuc m 2. All patients' sera showed similar ODs in ELISAs with natural and recombinant profilin. CONCLUSION: rCuc m 2 evoked strong immuno-reactivity equivalent to nCuc m 2, and has potential for diagnosis of melon allergy.
Entities:
Keywords:
Allergy; Cuc m 2; Melon; Natural allergen; Recombinant allergen
Authors: E Batanero; M A Gonzalez De La Peña; M Villalba; R I Monsalve; M Martin-Esteban; R Rodríguez Journal: Clin Exp Allergy Date: 1996-12 Impact factor: 5.018