抗尿激酶单克隆抗体识别相应抗原决定簇的研究

 尿激酶是一种纤溶酶原激活剂,临床上用于治疗血栓。为了有效地用单克隆抗体亲和柱纯化尿激酶,我们对一组抗尿激酶单克隆抗体识别相应抗原决定簇的特性进行了研究。Western Blotting试验表明:S_(13)、S_(26)、N_(14)、N_(30)、N_(17-2)、N_(34)、N_(36)七个单克隆抗体主要抗54000道尔顿的高分子量尿激酶(HUK)。除N_(30)外,其余抗体还同时不同程度地抗33000道尔顿的低分子量尿激酶(LUK)。N_(30)除识别HUK外,还识别分子量为18000道尔顿的多肽链。竞争性结合试验证明:七个单克隆抗体分别抗五个不同的抗原决定簇,但它们都不抗尿激酶的活力中心。

STUDY ON MONOCLONAL ANTIBODIES DEFINED ANTIGEN DETERMINANTS OF UROKINASE

Urokinase, a human plasminogen activator, is used clinically to promote the dissolution of thrombi.Availability of monoclonal antibodies against urokinase might provide a simple and efficient way to purify urokinase. With this in mind, we prepared seven monoclonal antibodies against urokinase using the hybridoma the-chnique. Western blotting experiments showed that the S13, S26, N14, N17, N30, N34, N36 antibodies recognized mainly the high molecular weight enzyme (54000 daltons, HUK) . Except for N30, these antibodies also reacted weakly to a shorter peptide chain (33000 daltons, LUK) . N30 recognized both HUK and the 18000 daltons fragment derived from the large molecule.Competitive binding assay demonstrated that these seven monoclonal antibodies might belong to five species, and recognized five different determinants respectively,but they were not able to inhibit the activity of urokinase.