title: A sensitive monoclonal antibody sandwich ELISA for the measurement of the major olive pollen allergen Ole e 1.
abstract: BACKGROUND
  Olive pollen is a major cause of inhalant allergy in countries around the Mediterranean sea. The major allergen of olive pollen is Ole e 1. Measurement of the major allergen content of allergen products for diagnosis and therapy is becoming an essential element of standardization protocols. This study aimed at the development of a monoclonal antibody (mAb) sandwich ELISA for Ole e 1.
  
  
  METHODS
  Balb/c mice immunized with Ole e 1 were used for the production of mAbs. Screening of mice and hybridomas was performed in a RIA with radiolabeled purified Ole e 1. Purified mAbs were used as catching and/or (biotinylated) detecting antibodies in sandwich ELISA.
  
  
  RESULTS
  Four mAbs (IgG1kappa) directed to nonoverlapping epitopes on Ole e 1 were obtained: 1A12, 5C1, 10A12 and 3H8. Both 1A12 and 10A12 were successfully used for affinity purification of Ole e 1 from olive pollen extract. Two sandwich ELISAs were developed, with 1A12 and 10A12 as catching, and 5C1 and 3H8 as detecting antibodies, respectively. Both catching and detecting antibodies were used in similar concentrations, ranging from 60 to 100 ng/well. For both ELISAs, the sensitivity was approximately 1 ng/ml of Ole e 1. The measuring range was from 1 to 25 ng/ml. No significant differences were observed, when the performance of both ELISAs in standardization of olive pollen extracts was compared.
  
  
  CONCLUSIONS
  Two sensitive sandwich ELISAs for the major olive pollen allergen Ole e 1 were developed. They will prove to be useful tools in allergen standardization protocols.
authors.affiliation: CLB and Laboratory for Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: van Ree
authors.forename: M
authors.initials: M
authors.lastname: Aalbers
authors.forename: O
authors.initials: O
authors.lastname: Kea
authors.forename: F M
authors.initials: FM
authors.lastname: Marco De La Calle
authors.forename: J M
authors.initials: JM
authors.lastname: Sempere Ortells
authors.forename: M
authors.initials: M
authors.lastname: Villalba
authors.forename: R
authors.initials: R
authors.lastname: Rodriguez
authors.forename: R C
authors.initials: RC
authors.lastname: Aalberse
mesh_terms: D000485:Allergens / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D051379:Mice; D008807:Mice, Inbred BALB C; D010940:Plant Proteins / Q000032:analysis*; D011058:Pollen; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C081946:Ole e I protein, Olea europaea; D010940:Plant Proteins