用分光光度法定量检测人血清巨噬细胞抑制因子1

Determination of Human Serum Macrophage Inhibitory Factor-1 by Spectrophotometry

结合基础酶联免疫吸附技术(ELISA),用分光光度法检测人血清中的巨噬细胞抑制因子1(MIC-1)的浓度。通过免疫反应将待测物质衍生化,生成可见光区可检测的复合物。在450nm波长处用分光光度法测定该复合物的吸光度,绘制MIC-1浓度-吸光度校准曲线,方程式为y=2.375-2.378/(1+0.001x)0.938(r=0.999)。根据样本的吸光度推算血清MIC-1的浓度。方法的线性范围为15.625—500.000pg/mL。回收率为98.2%—101.6%,相对标准偏差为1.89%—5.10%。

Combined with a sandwich enzyme-linked immunosorbent assay （ELISA）,the spectrophotometry method was applied to determine the serum concentration of human serum macrophage inhibitory factor-1（MIC／｜1）. After derivatization,the compounds which could be determined in visible region were generated. And the absorbance of them were measured by spectrophotometry at 450nm. Then a standard curve was constructed,with a four parameter logistic equation of y=2.375-2.378/（1＋0.001x）0.938 obtained（r=0.999）.Ultimately,the concentration of human serum MIC-1 were calculated by the standard curve. The best range for determination was 15.625—500.000pg/mL,with average recovery of 98.2%—101.6% and RSD of 1.89%—5.10%.