Development of ICT EIA to determine egg white allergen, L-PGDS aiming at reducing allergenicity.

Research Project

Project/Area Number	15K00833
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Eating habits
Research Institution	University of Kochi
Principal Investigator	Suzuki Makiko 高知県立大学, 健康栄養学部, 准教授 (60437001)
Co-Investigator(Kenkyū-buntansha)	田中守高知県立大学, 健康栄養学部, 助教 (00612350) 沼田聡高知県立大学, 健康栄養学部, 助教 (10565857)
Project Period (FY)	2015-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000) Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000) Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000) Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords	卵白アレルゲン / L-PGDS / ICT EIA / ICT EIA法 / rL-PGDS / 免疫複合体転移酵素免疫測定法
Outline of Final Research Achievements	We developed a sensitive Immune Complex Transfer Enzyme Immunoassay (ICT EIA) that enables us to measure levels of Egg white allergen, lipocalin-type prostaglandin D synthase (L-PGDS). The detection limit of this assay is 0.01 ng/ml, when we used recombinant L-PGDS as standards. This sensitivity is higher 2000-fold than ELISA which we have developed previously. However, we observed inhibitors against this assay in egg white. To identify this inhibitors, we analyzed L-PGDS complex. Our data shows that L-PGDS bind to ovalbumin. When we used 90% Ammonium sulfate fraction of egg white for determine L-PGDS in egg white by ICT EIA, the fluorescence intensity was dependent of egg white protein within 0.6～6 μg/ml.