摘要
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)标记的羊抗兔二抗与兔抗人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47.2%～101%;4℃有效期至少360d。结论成功建立双抗夹心ELISA检测BSP的方法。
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 polyclonal to BSP was used for detection, which was subseqently combined with goat anti - rabbit IgG - HRP. A sandwich ELISA method to detect BSP antigen 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 recovew 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.
出处
《广东医学》
CAS
CSCD
北大核心
2009年第11期1597-1599,共3页
Guangdong Medical Journal
基金
广东省自然科学基金重点项目(编号:06104396)
广东省医学科研基金项目(编号:B2007105)
