期刊
CHINESE SCIENCE BULLETIN
卷 58, 期 24, 页码 2938-2943出版社
SCIENCE PRESS
DOI: 10.1007/s11434-013-5702-9
关键词
immunoglobulin E; dot-blot assay; aptamer; biotin-avidin system
资金
- National Natural Science Foundation of China [30825027]
- Special Fund for Agro-Scientific Research in the Public Interest [200903009]
A novel aptamer based dot-blot assay for the detection of immunoglobulin E (IgE) was developed. A biotinylated aptamer was employed as the bio-recognition element to specifically interact with the target protein immobilized onto a nitrocellulose membrane substrate. Avidin conjugated horseradish peroxidase was introduced onto the membrane through the biotin-avidin system to catalyze the hydrogen peroxide mediated oxidation of 3,3',5,5'-tetramethylbenzidine, thereby producing the blue-colored insoluble product. The intensity of the dots increased as the concentration of IgE increased. The spot intensity was quantified using a simple portable instrument. A linear response relationship between the spot intensity and the concentration of IgE over the range of 50 nmol/L to 1 mu mol/L was obtained. The detection limit for IgE using the aptamer-based assay was 2.89 nmol/L. This assay was found to discriminate IgE from non-target proteins such as thrombin, bovine serum albumin and immunoglobulin G.
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