Develop an immunomagnetic bead based lateral flow assay for the rapid detection of a major peanut allergen – Ara h 2

• Main Authors: Yang-Fan Fung, 馮楊帆
• Other Authors: 溫曉薇
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/33010748556787855780

碩士 === 國立中興大學 === 食品暨應用生物科技學系所 === 105 === Peanut has been widely used in various kinds of food products, including snacks, biscuits, drinks and candies. However, peanut also induces severe allergic reactions, such as anaphylaxis shock. Thus, it is necessary to develop a rapid method for detecting peanut residues. Ara h2 is a major peanut allergen and its stability gets less impacted by food processing than another major peanut allergen- Ara h1. Therefore, in this study, we developed a magnetic bead labeled lateral flow assay to detect a major peanut allergen- Ara h2. Firstly, two anti-peptide antibodies and one anti-Ara h2 polyclonal antibody were used to determine an optimal pair of antibody: one antibody tagged on the surface of magnetic bead and the other immobilized on the test line of a lateral flow strip. Then, magnetic beads and NC membranes were optimizated. Following is the final condition, NC membrane type is AE99, testline antibody density is 7.5 μg/cmm magnetic beads size is 100 nm, modified antibody concentration is 400 μg/ml, NC membrane blocker is glycine, magnetic beads blocker is milk, ratio of magnetic beads and sample is 20:300, reaction time is 30 min. Furthermore, storability and inter/intra assay were tested. Therefore, a rapid magnetic bead labeled lateral flow assay was developed. LOD is 0.5 μg/ml. y = -0.1207x2 + 7.7389x + 2.0215, R² = 0.9997. Finally, this developed assay was used to detect Ara h 2 in oil product samples.