Development of new-type radiommunoassay system with higher sensitivity and lower radioactive waste disposal.

开发灵敏度更高、放射性废物处理更少的新型放射免疫分析系统。

基本信息

批准号：
04557049
负责人：
FUJIBAYASHI Yasuhisa
金额：
$ 4.22万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04557049/
关键词：
radioimmunoassay high specific activity metal label gallium Indium antisense oligonucleotide antibody digoxin 高感度ガリウム-67 銅-62

项目摘要

To achieve higher sensitivity in the detection of biologically active substances (ca..antibodies, drugs, antisense oligonucleotides, etc.) and lessen radioactive waste disposal, short-lived gamma emitting metal radionuclide was attached to such molecules. Gallium-67(Ga-67, half life=78hr)or indium-111 (In-111, half life=2.8day) was selected as a metalradionuclide and biologically active substances conjugated with deferoxamine, diethylenetriamine-pentaacetic acid or isothiocyanobenzyl-EDTA were radiolabeled.(1) Antibody : monoclonal anti-human growth hormone (GH) antibody was conjugated with deferoxiamine (DF) through dialdehyde-starch polymer spacer and labeled with Ga-67. Using this Ga-67-anti-GH antibody, immunoradiometric assay with 10 times higher sensitivity could be realized.(2) Drug : digoxin, a cardiac glycoside requireing therapeutic drug monitoring, was labeled with Ga-67 using DF as a bifunctional chelating agent. Ga-67-DF-digoxin has similar affinity to anti-digoxin antibody as well as Na, K-ATPase. Using the Ga-67-DF-digoxin, digoxin-RIA as well as myocardial Na, K-ATPase imaging in vivo.were successfully performed.3) Antisense oligonucleotide : Antisense oligonucleotide against human c-erbB-2 was 5'-aminated and conjugated with isothiocyanobenzyl-EDTA (IBE) and labeled with In-111. In-111-IBE-oligonucleotide showed high stability even in human serum, an indication of possible in-vivo application. In fact, biodistribution study in nude mice bearing c-erbB2 mRNA (+) tumor indicated the sequence selective tumor localization.It was concluded that short-lived metal radionuclide can be used not only for in vivo nuclear medicine, but in vitro diagnosis as well as basic research studies.

为了在检测生物活性物质（例如抗体、药物、反义寡核苷酸等）时获得更高的灵敏度并减少放射性废物处理，将短寿命的伽马发射金属放射性核素附着在此类分子上。选择镓67(Ga-67，半衰期=78小时)或铟-111(In-111，半衰期=2.8天)作为金属放射性核素，并与去铁胺、二乙烯三胺-五乙酸或异硫氰基苄基-EDTA缀合的生物活性物质(1) 抗体：单克隆抗人生长激素 (GH) 抗体与去铁胺结合(DF) 通过二醛-淀粉聚合物间隔基并用 Ga-67 标记。使用这种Ga-67-抗GH抗体，可以实现灵敏度提高10倍的免疫放射测定。(2)药物：地高辛，一种需要治疗药物监测的强心苷，使用DF作为双功能螯合剂用Ga-67标记。 Ga-67-DF-地高辛与抗地高辛抗体以及 Na,K-ATP 酶具有相似的亲和力。使用Ga-67-DF-地高辛、地高辛-RIA以及心肌Na、K-ATP酶体内成像均成功进行。3) 反义寡核苷酸：针对人c-erbB-2的反义寡核苷酸是5'-胺化的并且与异硫氰基苄基-EDTA (IBE) 缀合并用 In-111 标记。 In-111-IBE-寡核苷酸即使在人血清中也表现出高稳定性，这表明可能的体内应用。事实上，c-erbB2 mRNA (+)肿瘤裸鼠的生物分布研究表明了序列选择性肿瘤定位。得出的结论是，短寿命金属放射性核素不仅可用于体内核医学，还可用于体外诊断作为基础研究。