Ali Eman Ali, Nakamura Ryosuke, Falcone Franco H
Division of Molecular Therapeutics and Formulation, School of Pharmacy, University of Nottingham, Boots Science Bldg., Science Road, Nottingham, NG7 2RD, UK.
Division of Medicinal Safety Science, National Institute of Health Sciences, Setagaya-ku, Tokyo, Japan.
Methods Mol Biol. 2017;1592:147-161. doi: 10.1007/978-1-4939-6925-8_12.
Allergen-specific Immunoglobulin E (IgE) determination lies at the heart of diagnosis of sensitization to food and other allergens. In the past few years, reporter systems capable of detecting the presence of allergen-specific IgE have been developed by several labs. These rely on humanized rat basophil leukemia cell lines stably transfected with reporter genes such as firefly luciferase. In this chapter, we describe protocols for the use of the RS-ATL8 cell line (IgE cross-linking-induced luciferase expression; EXiLE) in 96-well and 384-well formats. We also describe optional treatment steps for enveloped virus and complement inactivation.
过敏原特异性免疫球蛋白E（IgE）的测定是食物和其他过敏原致敏诊断的核心。在过去几年中，几个实验室开发了能够检测过敏原特异性IgE存在的报告系统。这些系统依赖于稳定转染了诸如萤火虫荧光素酶等报告基因的人源化大鼠嗜碱性粒细胞白血病细胞系。在本章中，我们描述了在96孔板和384孔板形式中使用RS-ATL8细胞系（IgE交联诱导的荧光素酶表达；EXiLE）的方案。我们还描述了包膜病毒和补体灭活的可选处理步骤。
