title: Quantification in mass units of group 1 grass allergens by a monoclonal antibody-based sandwich ELISA.
abstract: BACKGROUND
  Grass pollen extracts currently used for allergy diagnosis and immunotherapy are a complex mixture of proteins of which only a few have allergenic activity. Lol p 1 is one of the most important allergens in grass pollen extracts.
  
  
  OBJECTIVE
  To develop a two-site enzyme-linked immunosorbent assay for the quantification of Lol p 1 and other group 1 allergens from grass species, and to assess its suitability for quantifying this group of allergens.
  
  
  METHODS
  Balb/c mice immunized with recombinant Lol p 1 were used for the production of monoclonal antibodies. Screening of hybridomas was performed by direct ELISA, and selected monoclonal antibodies were immobilized on ELISA plates and incubated with samples containing group 1 allergens. Bound allergens were detected by a combination of biotinylated Lol p 1-specific monoclonal antibody and peroxidase-streptavidin conjugate.
  
  
  RESULTS
  The assay is based on three Lol p 1-specific monoclonal antibodies with different epitope specificities. The optimized ELISA measured Lol p 1 concentrations ranging from 125 to 1000 ng/mL and could quantify group 1 allergen from grass species belonging to the Pooidea subfamily. The assay does not depend on anti-sera production or availability of human sera and thus reactives can be produced in unlimited amounts.
  
  
  CONCLUSIONS
  This sensitive and specific Lol p 1 assay will be helpful both for quantifying the group 1 allergen content of Pooideae pollen extracts intended for clinical use and for studying cross-reactivities among pollen extracts.
authors.affiliation: Research & Development Department, Bial-ARISTEGUI, Bilbao, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Arilla
authors.forename: I
authors.initials: I
authors.lastname: Ibarrola
authors.forename: E
authors.initials: E
authors.lastname: Eraso
authors.forename: M
authors.initials: M
authors.lastname: Aguirre
authors.forename: A
authors.initials: A
authors.lastname: Martínez
authors.forename: J A
authors.initials: JA
authors.lastname: Asturias
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism*; D052179:Antigens, Plant; D001667:Binding, Competitive / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008129:Lolium / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010940:Plant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D006109:Poaceae / Q000276:immunology*; D011058:Pollen / Q000235:genetics / Q000276:immunology*
chemical_list: D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C042814:Lol p I protein, Lolium perenne; D010940:Plant Proteins