title: Validity of using recombinant melon profilin, Cuc m 2, for diagnosis of melon allergy.
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.
  
  
  CONCLUSIONS
  rCuc m 2 evoked strong immuno-reactivity equivalent to nCuc m 2, and has potential for diagnosis of melon allergy.
authors.affiliation: Immunobiochemistry Lab, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.affiliation: Immunobiochemistry Lab, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Yaser
authors.initials: Y
authors.lastname: Bagheri
authors.affiliation: Razi Vaccine & Serum Research Institute, Mashhad, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Vahedi
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Farahzad
authors.initials: F
authors.lastname: Jabbari Azad
authors.affiliation: Immunobiochemistry Lab, Immunology Research Center, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Abdol-Reza
authors.initials: AR
authors.lastname: Varasteh
keywords: Allergy; Cuc m 2; Melon; Natural allergen; Recombinant allergen