Development of comprehensive allergenic epitope exploring method by means of EXiLE and mass spectrometry

2015 Fiscal Year Final Research Report

• Project/Area Number: 25460082
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: National Institute of Health Sciences
• Principal Investigator: NAKAMURA Ryosuke 国立医薬品食品衛生研究所, 医薬安全科学部, 室長 (50333357)
• Co-Investigator(Kenkyū-buntansha): NAKAMURA Rika 国立医薬品食品衛生研究所, 代謝生化学部, 研究員 (40643333)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: アレルゲン / EXiLE法 / 質量分析法 / IgE / ルシフェラーゼ / エピトープ / 抗原抗体反応

Outline of Final Research Achievements

In the present study, we aimed to develop a new allergenic epitope exploring method by means of the EXiLE (IgE crosslinking-induced luciferase expression) testing method. The EXiLE test is based on the luciferase assay using humanized rat basophilic leukemia cells (RS-ATL8) that express human high-affinity IgE receptor (FcεRI). E-C1 and E-G5 are murine monoclonal IgE antibodies against ovalbumin (OVA), with and without degranulation activity, respectively. RS-ATL8 cells sensitized with E-C1 were reactive to OVA in the liquid phase, but not in the case of E-G5. However, immobilized OVA reacted to both E-C1 and E-G5. We next tried to detect EXiLE responses to epitope peptide of E-C1. It was impossible to detect directly-immobilized epitope peptide with ELISA or EXiLE assay. However, when the peptide was conjugated to a carrier protein, the IgE response was observed, particularly with EXiLE assay.

Free Research Field

医歯薬学