Zhang Qianyun, Wang Xu, Li Zhenjia, Lin Jin-Ming
School of Science, Beijing University of Chemical Technology, Beijing 100029, China.
Anal Chim Acta. 2009 Jan 12;631(2):212-7. doi: 10.1016/j.aca.2008.10.042. Epub 2008 Nov 1.
In this work, the monoclonal antibodies (McAbs) to alpha-fetoprotein (AFP) were immobilized on two different solid phases, i.e., magnetic particles (MP) and coated tubes (CT). Based on this, a MP based chemiluminescence enzyme immunoassay (MP-CLEIA) and a CT based CLEIA (CT-CLEIA) were proposed for the evaluation of AFP in human serum and their analytical merits were studied and compared. By detailed discussion of several performance variants, including the concentration of immobilized McAb, dilution ratio of horseradish peroxidase (HRP) labeled McAb (HRP-McAb), total assay time, substrate volume, chemiluminescent kinetics, and hook effect concentration, the advantages of MP-CLEIA became conspicuously apparent. Moreover, in the presence of MP, the catalytic activity of labeled enzyme was kept to high extent and the stability of immunoreagents was satisfied. Finally, 59 human serum samples were detected by the MP-CLEIA and a good correlation was obtained when comparing the results with that from a commercial electrochemiluminescence immunoassay kit.
在本研究中，将抗甲胎蛋白（AFP）的单克隆抗体（McAbs）固定在两种不同的固相载体上，即磁性颗粒（MP）和包被管（CT）。基于此，提出了一种基于磁性颗粒的化学发光酶免疫分析法（MP-CLEIA）和一种基于包被管的化学发光酶免疫分析法（CT-CLEIA）用于检测人血清中的AFP，并对它们的分析性能进行了研究和比较。通过详细讨论几个性能变量，包括固定化单克隆抗体的浓度、辣根过氧化物酶（HRP）标记的单克隆抗体（HRP-McAb）的稀释比例、总分析时间、底物体积、化学发光动力学以及钩状效应浓度，MP-CLEIA的优势变得十分明显。此外，在有磁性颗粒存在的情况下，标记酶的催化活性能在很大程度上得以保持，免疫试剂的稳定性也令人满意。最后，用MP-CLEIA检测了59份人血清样本，将结果与商用电化学发光免疫分析试剂盒的结果进行比较时，获得了良好的相关性。
