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Standardization of Alternaria Alternata: Extraction and Quantification of Alt a 1 by Using an mAb-based 2-site Binding Assay

Standardization of Alternaria Alternata: Extraction and Quantification of Alt a 1 by Using an mAb-based 2-site Binding Assay

E Aden, B Weber, J Bossert, M Teppke, E Frank, R Wahl, H Fiebig, O Cromwell

J Allergy Clin Immunol. 1999 Jul;104(1):128-35.

PMID: 10400850

Abstract:

Background:
Alternaria alternata is recognized as an important cause of allergic disease. As with other molds, the extracts of A alternata used for diagnosis and therapy are highly heterogeneous, and there is a need for improved standardization. The major allergen Alt a 1 is well characterized and has been produced as a recombinant protein, but very few data are available on the Alt a 1 content in extracts.
Objective:
An assay for the quantification of Alt a 1 was developed and used for monitoring batch-to-batch consistency of A alternata extracts, and the correlation between skin prick test responses and Alt a 1 concentrations was studied.
Methods:
A 2-site binding assay based on an Alt a 1-specific mAb was developed and used for the quantification of Alt a 1 in allergen extracts. Quantitative skin prick tests were performed on 16 A alternata-sensitive patients and correlated with the Alt a 1 concentration.
Results:
The Alt a 1-specific mAb was found to be suitable for affinity purification, as well as for a 2-site binding quantification assay. In allergen extracts the Alt a 1 content was estimated as 2% to 4.7% of total protein. Quantitative skin prick tests showed an Alt a 1 concentration-dependent response.
Conclusion:
Quantification of Alt a 1 in A alternata extracts reflects their batch-to-batch consistency. Skin prick test responses to a standardized A alternata extract correlate with the Alt a 1 contents. An extract containing 3.7 microgram/mL Alt a 1 caused a response equal to that of a 1% histamine dihydrochloride solution.

Chemicals Related in the Paper:

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AS212294	N-Desmethylolanzapine-d8 dihydrochloride solution			Price

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