Comparison of multiplex immunoassay platforms.
Comparison of multiplex immunoassay platforms.
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DOI：
10.1373/clinchem.2009.135087
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发表时间：
2010-02
影响因子：
9.3
通讯作者：
Van Eyk JE
中科院分区：
文献类型：
--
作者：
Fu Q;Zhu J;Van Eyk JE
用高通量蛋白质组学技术发现的候选生物标志物（以及文献中报道的许多生物标志物）必须经过严格验证。对大群体中多种潜在生物标志物的同时定量评估对该领域提出了重大挑战。多重免疫测定是一种有前途的解决方案，有可能通过并行分析提供定量数据。与传统 ELISA 相比，这些测定方法所需的样品和试剂也少得多（传统 ELISA 因其仅测量单一抗原的能力而进一步受到限制）。我们测量了市售多重免疫测定的再现性、可靠性、稳健性、准确性和通量，以确定它们是否适合血清生物标志物分析和验证。检测平台 MULTI-ARRAY (Meso Scale Discovery)、Bio-Plex (Bio-Rad Laboratories)、A2 (Beckman Coulter)、FAST Quant (Whatman Schleicher & Schuell BioScience) 和 FlowCytomix (Bender MedSystems) 被选为目前用于高通量免疫分析的技术的代表性示例。所有测定均按照制造商指定的方案并使用其试剂盒中包含的试剂（稀释剂、校准剂、封闭剂和检测抗体混合物）进行。每个测定和抗原确定的可量化区间基于精度（CV < 25%）和回收百分比（测量浓度在实际浓度的 20% 以内）。 MULTI-ARRAY 和 Bio-Plex 检测具有最佳性能和最低检测限，并且 MULTI-ARRAY 系统在最宽的浓度范围（105 至 106）内具有最线性的信号输出。通过 MULTI-ARRAY 和 Bio-Plex 检测进一步研究了来自健康个体的未加标和细胞因子加标血清样品中的细胞因子浓度。 MULTI-ARRAY 和 Bio-Plex 多重免疫分析系统最适合生物标志物分析或定量。
Candidate biomarkers discovered with high-throughput proteomic techniques (along with many biomarkers reported in the literature) must be rigorously validated. The simultaneous quantitative assessment of multiple potential biomarkers across large cohorts presents a major challenge to the field. Multiplex immunoassays represent a promising solution, with the potential to provide quantitative data via parallel analyses. These assays also require substantially less sample and reagents than the traditional ELISA (which is further limited by its ability to measure only a single antigen). We have measured the reproducibility, reliability, robustness, accuracy, and throughput of commercially available multiplex immunoassays to ascertain their suitability for serum biomarker analysis and validation. Assay platforms MULTI-ARRAY (Meso Scale Discovery), Bio-Plex (Bio-Rad Laboratories), A2 (Beckman Coulter), FAST Quant (Whatman Schleicher & Schuell BioScience), and FlowCytomix (Bender MedSystems) were selected as representative examples of technologies currently used for high-throughput immunoanalysis. All assays were performed according to protocols specified by the manufacturers and with the reagents (diluents, calibrators, blocking reagents, and detecting-antibody mixtures) included with their kits. The quantifiable interval determined for each assay and antigen was based on precision (CV < 25%) and percentage recovery (measured concentration within 20% of the actual concentration). The MULTI-ARRAY and Bio-Plex assays had the best performance with the lowest limits of detection, and the MULTI-ARRAY system had the most linear signal output over the widest concentration range (105 to 106). Cytokine concentrations in unspiked and cytokine-spiked serum samples from healthy individuals were further investigated with the MULTI-ARRAY and Bio-Plex assays. The MULTI-ARRAY and Bio-Plex multiplex immunoassay systems are the most suitable for biomarker analysis or quantification.
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影响因子：
37.8
作者：
Sabatine, MS;Morrow, DA;Braunwald, E
通讯作者：
Braunwald, E
影响因子：
2.6
作者：
Utz, P
通讯作者：
Utz, P
影响因子：
7
作者：
Kingsmore, Stephen F.;Kennedy, Neil;Edgar, J. David M.
通讯作者：
Edgar, J. David M.
影响因子：
46.9
作者：
Schweitzer, B;Roberts, S;Kingsmore, SF
通讯作者：
Kingsmore, SF
影响因子：
7
作者：
Zolg, JW;Langen, H
通讯作者：
Langen, H