An avidin-biotin microELISA for rapid measurement of total and allergen-specific human IgE

Rao, PVS and Mccartneyfrancis, NL and Metcalfe, DD (1983) An avidin-biotin microELISA for rapid measurement of total and allergen-specific human IgE. In: Journal of Immunological Methods, 57 (1-3). pp. 71-85.

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Abstract

his paper describes an improved microtiter solid-phase enzyme immunoassay for the determination of total and allergen-specific human IgE. This assay technique is unique in its use of the avidin-biotin interaction to increase sensitivity. The avidin-biotin microtiter enzyme-linked immunosorbant assay (AB-microELISA) was performed in polyvinyl chloride microtiter plates using biotinylated anti-IgE and horseradish peroxidase (HRP)-avidin conjugate. This AB-microELISA technique enabled the quantitation of human serum IgE in the range of 0.1–5 ng/ml (10–500 pg/test) in less than 3 h. Total serum IgE, whether measured by the AB-microELISA or the paper radioimmunosorbant test (PRIST) was similar (correlation coefficient, r = 0.92). Further, the presence or absence of positive skin tests to 7 specific allergens determined in serum donors generally agreed with the presence or absence of allergen-specific IgE in their sera as measured by the AB-microELISA. The quantity of short ragweed allergen-specific IgE as determined by the AB-microELISA agreed with values obtained by the radioimmunosorbant test (RAST) (correlation coefficient, r = 0.89). No significant interference by ragweed-specific IgG (blocking antibody) was observed in the quantitation of allergen-specific IgE. The AB-microELISA is not only rapid and inexpensive, but also more sensitive than other published ELISA procedures and comparable to solid-phase radioimmunoassays in the quantitation of total and allergen-specific IgE.

Item Type:	Journal Article
Publication:	Journal of Immunological Methods
Publisher:	Elsevier Science
Additional Information:	Copyright of this article belongs to Elsevier Science.
Keywords:	IgE;ELISA;allergy
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	01 Jul 2009 09:48
Last Modified:	19 Sep 2010 05:35
URI:	http://eprints.iisc.ac.in/id/eprint/20910

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