Preparation of Recombinant Milk Allergen β-Lactoglobulin and Establishment of Magnetic Particle Chemiluminescence Detection Method

doi:10.13560/j.cnki.biotech.bull.1985.2019-0661

Abstract

Abstract:

Using prokaryotic system to express milk β-lactoglobulin（Bos d5）protein,a magnetic nanoparticle chemiluminescence immunoassay method was established to detect specific Ig E antibody content of allergen milk β-lactoglobulin. The recombinant bovine β-lactoglobulin gene was artificially optimized and synthesized with the plasmid,which then was introduced into the prokaryotic expression strain Rosetta to express the target protein. The purified recombinant Bos d5 protein was biotinylated and used on chemiluminescence platform,and the allergen items were detected on the chemiluminescence platform. The 22.5 kD recombinant bovine β-lactoglobulin with >85% purity was obtained and the biotinylated recombinant protein was used to develop the bovine β-lactoglobulin magnetic nanoparticle detection kit. The positive coincidence rate of serum samples was 88.9%,the negative coincidence rate was 97.3%,the total coincidence rate was 94/5% P<0.001,χ²=84.238,Kappa=0.874,while aligned with Phadia reference kit in 110 serum samples. The recombinant milk allergy component β-lactoglobulin obtained from the prokaryotic expression system has high immunoactivity,and the developed immunoassay kit performs well and can be used in medical clinical diagnosis.

Key words: milk, β-Lactoglobulin, allergens, recombinant protein, magnetic nanoparticle chemiluminescence immunoassay

HANG Xin-ru, GENG Rong-qing, QIAN Lin. Preparation of Recombinant Milk Allergen β-Lactoglobulin and Establishment of Magnetic Particle Chemiluminescence Detection Method[J]. Biotechnology Bulletin, 2020, 36(3): 193-198.

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