National-Reginoal Key Technology Engineering Laboratory for Medical Ultrasound, Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, Department of Biomedical Engineering, Medical School, Shenzhen University, Guangdong, 518020, People's Republic of China.
Luminescence. 2013 Nov-Dec;28(6):927-32. doi: 10.1002/bio.2461. Epub 2013 Jan 15.
Protein S100B is a clinically useful non-invasive biomarker for brain cell damage. A rapid chemiluminescence immunoassay (CLIA) for S100B in human serum has been developed. Fluorescein isothiocyanate (FITC) and N-(aminobutyl)-N-(ethylisoluminol) (ABEI) are used to label two different monoclonal antibodies of anti-S100B. Protein S100B in serum combines with labeled antibodies and can form a sandwiched immunoreaction. A simplified separation procedure based on the use of magnetic particles (MPs) that were coated with anti-FITC antibody is performed to remove the unwanted materials. After adding the substrate solution, the relative light unit (RLU) of ABEI is measured and is found to be directly proportional to the concentration of S100B in serum. The relevant variables involved in the CLIA signals are optimized and the parameters of the proposed method are evaluated. The results demonstrate that the method is linear to 25 ng/mL S100B with a detection limit of 0.02 ng/mL. The coefficient of variation (CV) is < 5% and < 6% for intra- and interassay precision, respectively. The average recoveries are between 97 and 107%. The linearity-dilution effect produces a linear correlation coefficient of 0.9988. Compared with the commercial kit, the proposed method shows a correlation of 0.9897. The proposed method displays acceptable performance for quantification of S100B and is appropriate for use in clinical diagnosis.
S100B 蛋白是一种临床上有用的非侵入性脑损伤生物标志物。现已开发出用于人血清中 S100B 的快速化学发光免疫分析（CLIA）。荧光素异硫氰酸酯（FITC）和 N-（氨基丁基）-N-（乙基异鲁米诺）（ABEI）用于标记抗 S100B 的两种不同单克隆抗体。血清中的 S100B 与标记的抗体结合，可形成夹心法免疫反应。基于使用包被有抗 FITC 抗体的磁性颗粒（MPs）进行简化的分离程序，以去除不需要的材料。加入底物溶液后，测量 ABEI 的相对光单位（RLU），发现其与血清中 S100B 的浓度成正比。优化 CLIA 信号涉及的相关变量，并评估所提出方法的参数。结果表明，该方法在 25ng/mL S100B 范围内呈线性，检测限为 0.02ng/mL。内和日间精密度的 CV 分别小于 5%和 6%。平均回收率在 97%至 107%之间。线性稀释效应产生的线性相关系数为 0.9988。与商业试剂盒相比，该方法的相关性为 0.9897。该方法在 S100B 的定量方面表现出可接受的性能，适用于临床诊断。
