title: Standardization of allergen products: 3. Validation of candidate European Pharmacopoeia standard methods for quantification of major birch allergen Bet v 1.
abstract: The BSP090 project aims at establishing European Pharmacopoeia Reference Substances in combination with the corresponding ELISA methods for the quantification of major allergens in allergen products. Two sandwich ELISAs proved suitable for quantification of Bet v 1, the major birch pollen allergen, in preceding phases of BSP090.
  
  
  
  Two Bet v 1-specific ELISA systems were compared with respect to accuracy and precision in a ring trial including 13 laboratories. Model samples containing recombinant rBet v 1.0101 as well as native birch pollen extracts were measured independently at least three times in each facility. The assessment was completed with a comparative quantification of Bet v 1 in 30 marketed birch allergen products in one laboratory, simulating the future use as reference method.
  
  
  
  In the collaborative study, both candidate ELISAs confirmed their suitability to quantify recombinant and native Bet v 1. ELISA-A showed higher precision and lower interlaboratory variability, yet ELISA-B exhibited slightly higher accuracy. Subsequent parallel measurement of Bet v 1 in a panel of 'real-life' birch allergen products indicated better repeatability of ELISA-B. Both systems detected substantial differences in Bet v 1 content between allergen products, but the effect was more pronounced using ELISA-B due to persistently higher values compared to ELISA-A.
  
  
  
  In the collaborative study, no deciding differences were observed between the two candidate ELISAs. Further comparison under conditions simulating the intended use combined with the criterion of long-term availability enabled the selection of one Bet v 1-specific ELISA for proposal as European Pharmacopoeia standard method.
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kaul
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Zimmer
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Dehus
authors.affiliation: European Directorate for the Quality of Medicines & HealthCare (EDQM), Strasbourg, France.
authors.forename: A
authors.initials: A
authors.lastname: Costanzo
authors.affiliation: European Directorate for the Quality of Medicines & HealthCare (EDQM), Strasbourg, France.
authors.forename: A
authors.initials: A
authors.lastname: Daas
authors.affiliation: European Directorate for the Quality of Medicines & HealthCare (EDQM), Strasbourg, France.
authors.forename: K H
authors.initials: KH
authors.lastname: Buchheit
authors.affiliation: BIAL Industrial Farmacéutica, Zamudio, Spain.
authors.forename: J A
authors.initials: JA
authors.lastname: Asturias
authors.affiliation: ALK-Abelló S/A, Madrid, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Barber
authors.affiliation: Laboratorios Leti S.L., Tres Cantos, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Carnés
authors.affiliation: Indoor Biotechnologies, Charlottesville, VA, USA.
authors.forename: M
authors.initials: M
authors.lastname: Chapman
authors.affiliation: ANSM, Saint Denis Cedex, France.
authors.forename: J
authors.initials: J
authors.lastname: Dayan-Kenigsberg
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Döring
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Führer
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: K M
authors.initials: KM
authors.lastname: Hanschmann
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Holzhauser
authors.affiliation: ALK-Abelló S/A, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Ledesma
authors.affiliation: Stallergenes Greer, Antony Cedex, France.
authors.forename: P
authors.initials: P
authors.lastname: Moingeon
authors.affiliation: Stallergenes Greer, Antony Cedex, France.
authors.forename: E
authors.initials: E
authors.lastname: Nony
authors.affiliation: ISS CRIVIB, Roma, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Pini
authors.affiliation: ALK-Abelló Inc., Round Rock, TX, USA.
authors.forename: G
authors.initials: G
authors.lastname: Plunkett
authors.affiliation: Allergopharma GmbH & Co. KG, Reinbek, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Reese
authors.affiliation: Danish Medicines Agency, Copenhagen S, Denmark.
authors.forename: E
authors.initials: E
authors.lastname: Sandberg
authors.affiliation: Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Ruhr-Universität Bochum (IPA), Bochum, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Sander
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Strecker
authors.affiliation: Food and Drug Administration, Silver Spring, MD, USA.
authors.forename: C
authors.initials: C
authors.lastname: Valerio
authors.affiliation: Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: van Ree
authors.affiliation: Paul-Ehrlich-Institut, Langen, Germany. stefan.vieths@pei.de.
authors.forename: S
authors.initials: S
authors.lastname: Vieths
mesh_terms: D000485:Allergens / Q000276:immunology; D052179:Antigens, Plant / Q000276:immunology; D029662:Betula / Q000276:immunology; D001688:Biological Products / Q000592:standards*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015203:Reproducibility of Results
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D001688:Biological Products
keywords: Bet v 1; ELISA; European Pharmacopoeia; biological standardisation; major allergen