Tamm-Horsfall糖蛋白放免分析测定盒的研制

A RADIOIMMUNOASSAY KIT FOR TAMM-HORSFALL GLYCOPROTEIN

采用氯化钠沉淀及SephadexG-100层析方法,从正常人尿中提纯了Tamm-Horsfall糖蛋白(THP)。用提纯的THP免疫家免制备了THP抗血清,抗体工作液滴度1:20000至1:40 000,亲和常数0.99×10^(10)l·mol^1,与人β_2-mG、白蛋白和IgG交叉反应分别小于0.02%,0.0001%和0.0004%。并采用双抗加PEG的方法制备了THP放免分析测定盒,对人血清和尿中THP进行测定。测定范围50—2000ng/ml,批内及批间变异系数分别为(1.6-2.9)%(n=6)和(2.3-4.2)%(n=6),回收率(96.2—103.9)%,正常人血清THP为(123.3±21.9)ng/ml(n=14),24小时尿THP排泄量为(29.0-43.9)mg(n=3)。

Tamm-Horsfall glycoprotein (THP) was prepared and purefied from urine by salt precipitation and chromatography on Sephadex G-100. An antiserum was raised by immunizing rabbits. The antiserum had a working titer of 1: 20 000 to 1 : 40 000 and the affinity constant was 0.99×10^(10)l. mol^(-1). The cross reaction with β_2-MG, HSA and IgG was lower than 0.02%, 0.000 1% and 0.000 4% respectively. A RIA kit was developed for determination of THP in human serum and urine. The standard range was 50—200 0 ng/ml and recoveries varied from 96.2% to 103.9%. Intra-and Inter-assay coefficient of variation were (1.6—2.9)% and (2.3—4.2)% respectively. The normal range of THP was 123. 3±21.9 ng/ml(X±SD, n=14) in serum, and (29.0—43.9) mg/ 24 h (n=3)in urine.