Protocol

Food Allergens

Volume 1592 of the series Methods in Molecular Biology pp 147-161

Date:

Use of Humanized RS-ATL8 Reporter System for Detection of Allergen-Specific IgE Sensitization in Human Food Allergy

  • Eman Ali AliAffiliated withDivision of Molecular Therapeutics and Formulation, School of Pharmacy, University of Nottingham
  • , Ryosuke NakamuraAffiliated withDivision of Medicinal Safety Science, National Institute of Health Sciences
  • , Franco H. FalconeAffiliated withDivision of Molecular Therapeutics and Formulation, School of Pharmacy, University of Nottingham Email author 

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Abstract

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.

Key words

Reporter system Luciferase Humanized RBL IgE Allergy RS-ATL8 EXiLE