Shima H, Saeki K, Yamanouchi M, Omori K, Sugiura M
Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Company Limited, Osaka, Japan.
Biochem Mol Med. 1995 Jun;55(1):61-5. doi: 10.1006/bmme.1995.1032.
The development of a sensitive enzyme immunoassay for endothelin is described. This assay is based on a sandwich method using two different monoclonal antibodies against endothelin-1. A monoclonal antibody, which reacted to the C-terminal region of endothelin, was selected as an immobilized antibody. The Fab' fragment of another monoclonal antibody, which might recognize the N-terminal rigid region of endothelin, was used as a horseradish peroxidase-labeled detector antibody. The assay measures endothelin-1 and endothelin-2 with a sensitivity of 1 fmol/ml. We have determined that cultured endothelial cells actually produced endothelin in significant amounts in a time-dependent manner. The levels of plasma endothelin extracted with Sep-Pak tC18 light cartridges could also be monitored. A basal endothelin level was about 0.3 fmol/ml of plasma, and a transient increase was observed 4 h after starting blood collection under in vivo experimentation in the rat. This enzyme immunoassay will facilitate the investigation of physiological roles of endothelin.
本文描述了一种用于内皮素的灵敏酶免疫测定法的开发。该测定法基于夹心方法，使用两种针对内皮素-1的不同单克隆抗体。一种与内皮素C末端区域反应的单克隆抗体被选作固定化抗体。另一种单克隆抗体的Fab'片段，其可能识别内皮素的N末端刚性区域，被用作辣根过氧化物酶标记的检测抗体。该测定法测量内皮素-1和内皮素-2的灵敏度为1 fmol/ml。我们已经确定，培养的内皮细胞实际上以时间依赖性方式大量产生内皮素。用Sep-Pak tC18轻型柱提取的血浆内皮素水平也可以进行监测。基础内皮素水平约为0.3 fmol/ml血浆，在大鼠体内实验中开始采血4小时后观察到短暂升高。这种酶免疫测定法将有助于研究内皮素的生理作用。
