双抗夹心ELISA法检测骨唾液酸蛋白的初步建立

目的 建立快速检测骨唾液酸蛋白的酶联免疫吸附试验（ELISA）检测方法，为试剂盒的研制奠定基础。方法 用鼠抗人骨唾液酸蛋白（BSP）单克隆抗体包被酶标板，兔抗人BSP检测，辣根过氧化物酶（HRP）标记的羊抗鼠二抗与兔抗人BSP相结合，初步建立BSP的双抗夹心ELISA检测方法。结果 自建双抗夹心ELISA法检测骨唾液酸蛋白的灵敏度为1.5ng/ml，标准曲线的线性范围为2-100ng/ml，回归方程y=0.2548x+0.0775 (r=0.992)；批内批间变异系数分别为4.6％-5.4％和5.2％-7.1％，对人血清做50、25和10ng/ml三个加标浓度的回收率实验，回收率在47.2％-101％之间；4℃有效期至少360天。结论：成功建立双抗夹心ELISA检测BSP的方法。

Establishment of a sandwich ELISA for quantitative measurement of bone sialoprotein

Objective To establish a sandwich ELISA for quantitative measurement of bone sialoprotein (BSP). Methods Anti -BSP monoclonal antibody (mAb) was coated on solid plate. Rabbit polyelonal to BSP was used for de-tection, which was subseqently combined with goat anti -rabbit lgG -HRP. A sandwich ELISA method to detect BSP an-tigen was established. Results The sensitivity of this assay was 1. 5 ng/mL, linear range was 2 ～ 100 ng/ml, The linear equation was y = 0. 254 8x + 0. 077 5 (r = 0. 992). The coefficients of variation were 4.6％ to 5.4％ within assay and 5. 2％ to 7. 1％ between assays. The recovery rate was 47.2％ to 101％. The antibody can be stored at 4℃ in 360 days at least. Conclusion A sandwich ELISA assay for detecting BSP is successfully established.