We recently bred a cucumisin-containing Korean variety of melon. However, it is not known whether the cucumisin macromolecule can pass the intestinal barrier. We developed a polyclonal antibody against cucumisin and tried to confirm whether orally administered cucumisin passes through the intestinal tract with mouse. Cucumisin was isolated and purified from the improved Korean melon by ion exchange chromatography. Protein size was estimated by SDS-PAGE and protease activity was confirmed by performing zymography and fibrin plate assay. The purified cucumisin largely comprised a native form (67 kDa) and mature form (54 kDa), and displayed fibrinolytic activity, as evident by the decomposition of gelatin and fibrin to form a clear zone. To confirm the transition to blood following the oral administration of cucumisin, a polyclonal antibody against cucumisin was produced. Use of the antibody in ELISA and immunoblotting analyses confirmed the transfer of cucumisin to the blood of cucumisin-fed mice as compared to those not fed cucumisin. Orally administered cucumisin can migrate through the intestinal barrier and into the blood. The fibrinolytic activity of cucumisin purified from the improved Korean melon might be exploited for development as a raw material for a thrombolytic drug.

中文翻译：

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抗来自大韩甜瓜的纯化黄瓜黄素的多克隆抗体及其应用：溶栓应用

最近，我们培育了一种含黄瓜黄素的韩国甜瓜品种。但是，尚不知道cucumisin大分子是否可以通过肠道屏障。我们开发了一种针对Cucumisin的多克隆抗体，并试图确认口服的Cucumisin是否与小鼠一起通过了肠道。通过离子交换色谱法从改良的朝鲜甜瓜中分离并纯化了黄瓜黄素。通过SDS-PAGE估计蛋白质大小，并通过进行酶谱和纤维蛋白平板测定法确认蛋白酶活性。纯化的cucumisin主要包含天然形式（67 kDa）和成熟形式（54 kDa），并显示出纤溶活性，明胶和纤维蛋白分解形成透明区即可证明这一点。为了确认口服黄瓜黄素后向血液的转化，产生了一种针对cucumisin的多克隆抗体。抗体在ELISA和免疫印迹分析中的使用证实了与未饲喂Cucumisin的小鼠相比，Cucumisin已转移至Cucumisin喂养的小鼠血液中。口服施用的cucumisin可以通过肠屏障迁移并进入血液。从改良的韩国甜瓜中提纯的姜黄素的纤溶活性可能会被开发用作溶栓药的原料。