A sensitive ELISA for the detection of bovine crude heparin in porcine heparin
A sensitive ELISA for the detection of bovine crude heparin in porcine heparin
复制标题
DOI：
10.1081/ias-100107398
复制
发表时间：
2001-11-01
影响因子：
--
通讯作者：
Levieux, D
中科院分区：
文献类型：
--
作者：
Levieux, A;Rivera, V;Levieux, D
肝素是一种有效的抗凝血药，几十年来一直是从牛或猪的组织中提纯出来的。然而，随着疯牛病的出现，肝素的纯化目前仅限于猪肠黏膜。为了控制粗肝素的来源，制备了针对牛污染物的多克隆抗体。这些抗体被用来制备一个三明治和两个竞争性间接ELISA。以牛粗肝素为包被物，抗牛粗肝素为检测抗体，建立了竞争性间接酶联免疫吸附试验。该方法的检出限为猪肝素中牛粗肝素1ppm。考虑到已知来源的10种粗猪肝素和9种纯猪肝素的背景可变性，检测水平为5ppm。绵羊和山羊粗肝素有轻微的交叉反应(6-17ppm)。
Heparin is an effective anticoagulant drug which has been purified for decades from bovine or porcine tissues. However, with the emergence of BSE, heparin purification is today restricted to porcine intestinal mucosa. To control the origin of crude heparins, polyclonal antibodies were raised against bovine contaminants. These antibodies were used to develop one sandwich and two competitive indirect ELISAs. Optimal results were obtained with competitive indirect ELISA, using bovine crude heparin for the coating and anti-bovine crude heparin as a detector antibody. The detection limit of the assay was I ppm of bovine crude heparin in a porcine heparin. Taking into account the variability of the background obtained for 10 crude and 9 pure porcine heparins from known origin, the detection level was 5 ppm. Ovine and caprine crude heparins cross-reacted slightly (6-17 ppm).