| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Outline of Final Research Achievements |
The purpose of this study is the production of novel antigen molecule for acquisition of “binder antibodies” to facilitate crystallization of G protein coupled receptor (GPCR), and for acquisition of “functional antibodies” to regulate the activity of GPCR. Firstly, we have improved the expression and purification system using budding yeast for producing the stable variant of a high-affinity type histamine receptor. The purified receptor variant was immobilized on the affinity beads via its C-terminal His-tag. Then the beads were covered by a lipid bilayer consisting of lipid and adjuvant, which gave “antigen particle”. The antigen particle and a proteoliposome containing the receptor were injected into mice for immunization, and the antibody titer for the receptor was measuredusing ELISA experiment. The ELISA showed that the serum for the antigen particle contained a higher ratio of the antibodies that recognize extracellular domain of the receptor.
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