Establishment of Immunoassay Method for Allergy-Inducing Proteins in Eggs

doi:10.7506/spkx1002-6630-201306029

Abstract

Abstract: Allergy-inducing proteins in eggs were isolated and purified by ammonium sulfate precipitation and ion exchange chromatography. Polyclonal antibody was generated by inoculated rabbits. A rapid, sensitive and specific ELISA method was developed and optimized for the detection of egg allergy proteins. Under the optimal experimental conditions, a good linear relationship between allergy-inducing proteins and inhibitory rate in the range of 1 × 10-4–10 μg/mL was observed. The regression equation was y = 0.74145＋0.21692 × lgC (r = 0.9971). IC50 and IC10 were 77.076 ng/mL and 1.104 ng/mL, respectively. The recovery rate of the ELISA method was 70.1%－117.8%. The developed method had cross-reactivity with the proteins from turkey, duck and goose eggs, despite having no cross-reactivity with the proteins from peanut or milk. The variation coefficients of intraassay and inter-assay were 4.63% and 5.49%, respectively. The shelf life of the prepared immunoassay kit was 6 months during the storage at 4 ℃ and met the requirements for the analysis of egg allergy proteins with fast operation and high sensitivity.

Key words: egg allergy proteins, enzyme linked immunosorbent assay, polyclonal antibody

CLC Number:

R155.5

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