Development of quartz-crystal-microbalance-based immunosensor array for clinical immuno pheno typing of acute leukemias
Development of quartz-crystal-microbalance-based immunosensor array for clinical immuno pheno typing of acute leukemias
复制标题
DOI：
10.1016/j.ab.2005.12.006
复制
发表时间：
2006-04-01
影响因子：
2.9
通讯作者：
Shen, GL
中科院分区：
文献类型：
--
作者：
Zeng, H;Wang, H;Shen, GL
已开发出一种集成压电免疫传感器阵列，用于临床急性白血病的免疫表型分析。每个石英晶体微天平 (QCM) 均采用正丁胺、纳米金颗粒和蛋白 A (PA) 的等离子体聚合膜制成，用于定向固定抗体。白血病谱系相关单克隆抗体分别固定在纳米金 PA 修饰的晶体表面上，该晶体由 2 x 2 型探针构成，形成基于 QCM 的免疫传感器阵列。考察了主要检测条件，包括抗体固定量、pH、免疫反应时间、样品稀释倍数等。通过免疫传感器阵列法、免疫组织化学和流式细胞术同时分析Jurkat细胞，探讨新技术免疫表型分型的可行性。研究发现，所开发的技术可以在 5 分钟内轻松识别白血病样本，并可以动态监测免疫反应过程。此外，使用基于QCM的免疫传感器方法和荧光免疫分析法对从96例急性白血病患者和24例正常受试者分离的有核细胞上表达的CID抗原进行了比较研究。采用基于免疫传感器的方法对患者样本进行免疫表型分型，768组数值数据的分型率为88.93%，经chi(2)分析，两种方法之间无显着统计学差异（chi(2) = 3.4，p > 0.05）。这种新型免疫传感器阵列具有灵敏度高、特异性高、重现性好、操作简便、成本低等优点。标本评估结果表明，其在临床上可能适用于人类分化白细胞的定量和急性白血病的免疫表型分析。 (c) 2005 Elsevier Inc. 保留所有权利。
An integrated piezoelectric immunosensor array has been developed to immunophenotype acute leukemias in clinic. Each quaftz crystal microbalance (QCM) was fabricated with plasma-polymerized film of n-butylamine, nanogold particles, and protein A (PA) to be used to immobilize antibodies in orientation. Leukemic lineage-associated monoclonal antibodies were separately immobilized onto the nanogold-PA-modified surface of the crystals, which were constructed by a 2 x 2 type of probes forming a QCM-based immunosensor array. The main detection conditions were investigated, including the immobilization amount of antibodies, pH, immunoreaction time, sample dilution ratio, etc. The immunophenotyping feasibility of the new technique was investigated through simultaneously analyzing Jurkat cells by the immunosensor array method, inimunohistochemistry, and flow cytometry. It was found that the developed technique Could readily identify leukemia samples in 5 min and might monitor dynamically the immunoreaction processes. Moreover, comparison studies were carried out for CID antigens expressed on the nucleated cells isolated from 96 acute leukemic patients and 24 normal subjects using the QCM-based immunosensor method and the fluoroimmunoassay. Results obtained by immunophenotyping patients' samples with the immunosensor-based method achieved the rate of 88.93% in 768 groups of numerical data, where no significant statistical difference was observed between the two methods when checked by chi(2) analysis (chi(2) = 3.4, p > 0.05). This new immunosensor array showed the merits of high sensitivity, high specificity, good reproducibility, easy operation, and low cost. The results of specimen evaluation indicated that it might be clinically suitable for quantifying human differentiated leukocytes and immunophenotyping of acute leukemias. (c) 2005 Elsevier Inc. All rights reserved.