成果報告書詳細
管理番号20140000000450
タイトル*平成25年度中間年報 「がん超早期診断・治療機器の総合研究開発 超早期高精度診断システムの研究開発:血液中のがん分子・遺伝子診断を実現するための技術・システムの研究開発 血中分子・遺伝子診断のための基礎技術の研究開発(がんの発症を予測するシステムの開発 小型診断用質量分析装置の開発によるエクソソーム診断)」
公開日2014/7/26
報告書年度2013 - 2013
委託先名塩野義製薬株式会社
プロジェクト番号P10003
部署名バイオテクノロジー・医療技術部
和文要約
英文要約[Establishment of Monoclonal Antibody against CD147]
To establish of the monoclonal antibodies against CD147, five A/J and five Balb/c mice were immunized 5 times every 3 weeks with an ectodomain of CD147 protein. After 5th immunization, two mice that show the highest antibody titers were selected for boost-immunization. However, animals were died after boost-immunization be-cause of Anaphylactic Shock. Then, immunization to establish of the monoclonal antibodies against CD147 has been re-started using ten A/J mice. After 5 times immunization, two mice that show the highest antibody titers were selected and boosted with ectodomain of CD147 of 1/10 amount in the normal use for boost- immunization. Three days after boost-immunization, spleen cells were fusion with the myeloma cells.
Currently, hybridomas are cloning by the limiting dilution technique.

[Establishment of Standard Protein]
 It is necessary for a stable standard to develop ELISA and ExoScreen system detecting CD147/CD9 double positive extracellular microvesicles as an in vitro diagnostic. Usually, Extracellular microvesicles have a poor stability for a freeze thawing and a long-term storage. Then, expression vector of an ectodomain of CD147 protein fused a large extracellular loop of CD9 protein was constructed and transfected to mammalian cells. Purified the fusion protein has a good activity in ELISA and ExoScreen system.
Currently, a stability of the fusion protein are examined.

[Confirmation of specificity]
No progress.
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