Research Paper ScFv化抗テネイシンC抗体による心疾患の分子標的イメージング診断法の確立

下條, 尚志  ,  SHIMOJO, Naoshi

2015-05-29 , 三重大学
テネイシン-C(TN-C)は虚血性心疾患などの炎症部位に一過性に高発現する。TN-Cを特異的に認識する抗体を用いて、分子診断薬として改良可能かどうかを検証した。ハイブリドーマのRNAから抗TN-C抗体のH鎖可変領域(VH)とL鎖可変領域(VL)遺伝子をクローニングし、重複伸長PCRを行い、VH-linker-VL遺伝子(scFv遺伝子)を大腸菌、in vitroセルフリー(RTS小麦胚芽)の発現系、哺乳類細胞株(HEK293)へそれぞれ遺伝子導入した。その結果、HEK293において最も多く発現量が得られた。準備が整い次第、心筋炎モデル動物に注入して病巣への集積を確認する予定である。
Tenascin-C (TN-C) is expressed transiently inflammatory regions in various cardiovascular diseases. We investigated the possibility that specific antibody against TN-C is useful for detecting cardiovascular diseases as a molecular diagnostic reagent. Heavy chain variable region (VH) and light chain variable region (VL) genes of anti-TN-C antibody were obtained from total RNA in hybridoma, and were performed overlap-extended PCR. Expression vector inserted VH-linker-VL gene (scFv) was used in E. coli, in vitro cell free (RTS wheat germ) and mammalian cell line (HEK293) expression systems. As a result, the antibody expression was highly effective in HEK293 cell line system. As soon as it’s ready, we are planning to experiments of antibody accumulation in foci of myocarditis model mice.

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