2017 Fiscal Year Final Research Report
Creation of the membrane-protein-particle antigen for the effective production of a crystallization-facilitator antibody
| Project/Area Number |
15K14460
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | Kyushu University |
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Principal Investigator |
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Keywords | Gタンパク質共役型受容体 / 抗原 / 膜タンパク質 |
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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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| Free Research Field |
タンパク質工学
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