可溶性尿激酶受体的免疫放射测定及其临床应用

Immunoradiometric assay of human soluble urokinase receptor and its clinical application

目的 以免疫放射测定法 (IRMA) ,特异性地检测可溶性尿激酶受体 (suPAR) ,以了解其生理水平及在病理状态下的改变。方法 应用 2株抗人suPAR的单克隆抗体 ,用氯胺T法以12 5I标记其中一株单抗 ,建立suPAR双抗夹心IRMA ,测定健康成人、良性肿瘤及恶性肿瘤患者血清suPAR的水平 ,并比较健康成人和阵发性睡眠性血红蛋白尿 (PNH)患者血浆suPAR的水平。结果 IRMA检测suPAR的最小可测值为 1 95 μg/L ,单抗与重组人uPAR的亲和常数为 4 75× 10 9mol/L ,平均回收率为1 0 13,批内及批间CV分别为 (6 40± 2 5 7) %和 (10 48± 2 6 5 ) %。 6 2名健康成人血清suPAR水平为(2 71± 1 12 ) μg/L ,2 1名健康成人血浆suPAR水平为 (1 73± 0 96 ) μg/L。与健康成人相比 ,2 4例良性肿瘤患者血清suPAR水平增高 ,而 47例恶性肿瘤患者血清suPAR水平则显著增高。PNH患者血浆suPAR水平也较健康成人增高。结论 suPAR以IRMA法检测敏感性、重复性好 ,准确性较高 ,恶性肿瘤患者和PNH患者血清 (浆 )

Objectives To establish a specific immunoradiometric assay (IRMA) for human soluble urokinase receptor (suPAR) and to investigate suPAR levels in normal adults and some pathological conditions. Methods Two monoclonal antibodies (McAb) were used, and one of them was labeled with 125 I by using chloramine T method to set up the IRMA for suPAR. Serum or plasma levels of normal adults, and patients with benign and malignant tumors and paroxysmal nocturnal hemoglobinuria (PNH) were measured. Results The detectable limit of this suPAR IRMA was 1 95 μg/L. The affinity constant was 4 75×10 9 L/mol. The mean rate of recovery was 101 3%, and the mean coefficient of variation for intra and interbatch assay were (6 40±2 57)% and (10 48±2 65)%, respectively. The mean levels of suPAR were(2 71±1 12) μg/L in serum of 62 normal adults and (1 73±0 96) μg/L in plasma of 21 normal adults, while those in serum of 24 benign tumor patients were (3 88±1 57) μg/L and (5 94±2 56) μg/L in serum of 47 malignant tumor patients. The mean plasma suPAR level of PNH patients was (4.06±2.73) μg/L. The suPAR levels of all these patients were higher than the normal levels. Conclusions The IRMA for suPAR is a sensitive, accurate and reproducible technique. Serum or plasma suPAR assay is useful in understanding the clinical status of tumor and PNH patients.