Objective 1: Pollen counts
Counting pollen with a Burkard pollen trap was established around 1960 and the method is standardized worldwide, well known, and often applied. All partners are very experienced in operating a Burkard pollen trap. No training is needed. Quality control will be done by sending each partner a set slides with known pollen counts (calibrating samples). In addition, 2 unknown samples will be sent to each partner yearly that need to be counted and send to the quality-controlling partner.
Objective 2: Allergen measurements
Measuring allergens in ambient air was established in the coordinators laboratory in 2002, and the method is commercially available worldwide. The way the assay is employed, ELISA, is a standard technique available since the early nineteen seventies. All partners in WP “allergens in ambient air” have experience with this method. Training is needed in order to achieve reproducible results between laboratories. The coordinator has 5 years experience with exactly the proposed methods. The ELISA is not complicated, but handling the pollen in a similar way will guarantee results that can be compared between laboratories. Quality control will be done with pre-determined and unknown samples to identify laboratories that are deviating from the standard. Quality control will be reinitiated each year. This will assure accurate and reproducible ELISA results.
Objective 3: Dissemination
The methods will be made available to all partners and public in the form of a publication at the end of the project.
This work package will provide the antibodies, standards, positive controls and quality control needed for the project. In brief the partner will deliver and distribute:
1. Capture and detection antibodies for the quantification of Bet v 1 and Phl p 5, produced by
Allergopharma, to be used for monoclonal antibody based 2-site binding assays, performed
as ELISA. The Ole e 1 antibodies are commercially available from Bial Aristegui in Spain.
2. Standards for the Bet v 1 and Phl p 5 ELISAs. Ole e 1 see above.
3. Generation of positive controls and unknowns for the assays. Ole e 1 see above.
4. Technical personnel will be trained for 1 week at Allergopharma in Reinbek with the aim of
achieving a sufficient inter-assay variability and consistent extraction of pollen samples.
5. Instructions (SOPs: Standard Operating Procedures), including quality control measures.
A “ring trial” will be performed to control inter-assay variability using samples which will be measured in each laboratory together with internal standards, followed by evaluation of the results by Allergopharma. Control of the results obtained from samples during the project will be done at Allergopharma by evaluation of the quality control samples.