A sensitive monoclonal antibody sandwich ELISA for the measurement of the major olive pollen allergen Ole e 1

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 diagno sis and therapy is becoming an essential element of standardization protoco ls. This study aimed at the development of a monoclonal antibody (mAb) sand wich ELISA for Ole e 1. Methods: Balb/c mice immunized with Ole e 1 were us ed for the production of mAbs. Screening of mice and hybridomas was perform ed in a RIA with radiolabeled purified Ore e 1. Purified mAbs were used as catching and/or (biotinylated) detecting antibodies in sandwich ELISA. Resu lts: Four mAbs (IgG1 kappa) directed to nonoverlapping epitopes on Ole el w ere obtained: 1A12, 5C1, 10A12 and 3H8. Both 1A12 and 10A12 were successful ly 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 a nd 3H8 as detecting antibodies, respectively. Both catching and detecting a ntibodies were used in similar concentrations, ranging from 60 to 100 ng/we ll. 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 fo r the major olive pollen allergen Ole e 1 were developed. They will prove t o be useful tools in allergen standardization protocols. Copyright (C) 2000 S. Karger AG, Basel.