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. 2012 Feb;67(2):235-41.
doi: 10.1111/j.1398-9995.2011.02750.x. Epub 2011 Nov 18.

A multi-allergen standard for the calibration of immunoassays: CREATE principles applied to eight purified allergens

S Filep  1 , A Tsay, L Vailes, G Gadermaier, F Ferreira, E Matsui, E M King, M D Chapman
Affiliations

A multi-allergen standard for the calibration of immunoassays: CREATE principles applied to eight purified allergens

S Filep et al. Allergy. 2012 Feb.

Abstract

Background: Allergen measurements are widely used for environmental exposure assessments and for determining the potency of allergen vaccines, yet few purified allergen standards have been developed. The aim of the study was to develop a single standard containing multiple purified allergens that could be used in enzyme immunoassays and in multiplex arrays for the standardization of allergen measurements.

Methods: Eight purified allergens were formulated into a single multi-allergen, or 'universal', standard based on amino acid analysis. Dose-response curves were compared with previous individual ELISA standards and allergen measurements of house dust extracts to obtain correction factors. Measured allergen concentrations were also modeled using linear regression, and the predictive accuracy was determined.

Results: Parallel dose-response curves were obtained between the universal allergen standard and the individual ELISA standards, with close agreement between curves for 5/8 allergens. Quantitative differences of greater than twofold were observed for Fel d 1, Can f 1, and Der f 1, which were confirmed by the analysis of house dust extracts. Correction factors were developed that allowed ELISA data to be expressed in terms of the universal standard. Linear regression data confirmed the predictive accuracy of the universal standard.

Conclusion: This study shows that a single standard of eight purified allergens can be used to compare allergen measurements by immunoassay. This approach will improve the continuity of environmental exposure assessments and provide improved standardization of allergy diagnostics and vaccines used for immunotherapy.

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Figures

Figure 1
Figure 1
SDS-PAGE analysis of purified allergens. Left to right: molecular weight markers, Der p 1, Der f 1, Der p 2, Fel d 1, Can f 1, Mus m 1, Rat n 1, Bla g 2.
Figure 2
Figure 2
Dose response curves of individual ELISA standards (■) and the UAS (▲) for eight allergens measured by ELISA.
See this image and copyright information in PMC

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