Protein microarray platform for the multiplex analysis of biomarkers in human sera
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- 作者：Teresa Urbanowska ; Sara Mangialaio ; Christophe Zickler ; Sirinda Cheevapruk ; Paul Hasler ; Stephan Regenass and Francois Legay
- 关键词：Biomarker ; Protein microarray ; Immunoassay
- 刊名：Journal of Immunological Methods
- 出版年：2006
- 出版时间：30 October 2006
- 年：2006
- 卷：316
- 期：1-2
- 页码：1-7
- 全文大小：287 K
文摘
Many biomarkers are currently used to monitor patients in clinical studies. Technologies which evaluate, validate and monitor biomarkers in a cost effective and efficient manner are a necessity. Here we describe the development, validation and implementation of a protein microarray platform for the quantitative and simultaneous analysis of six proteins: IL-1β, IL-1ra, IL-6, IL-8, MCP-1 and TNFα. The platform utilizes a 96-well plate as a solid support on which antibodies are immobilized using non-contact piezoelectric printing. The reaction is based on a sandwich ELISA and the signal is quantified by chemiluminescence with a CCD camera. The robustness and reproducibility of the methodology was investigated using the Food and Drug Administration (FDA) regulatory guidelines for pharmacokinetic assay validation, in which a spike-recovery validation test was elaborated and run over 3 days. The method was shown to be both quantitative and reproducible, with assay accuracy between 70 % and 130 % , and an assay precision of less than 30 % . In addition, protein microarray performance was compared with the classical ELISA approach. Sera collected from a total of 78 individuals were assayed using both approaches. Correlation coefficients (R2) between the two technologies were calculated for each of the analytes: 0.90 for IL-1β, 0.60 for IL-1ra, 0.93 for IL-6, 0.96 for IL-8, 0.94 for MCP-1 and 0.95 for TNFα. The results obtained demonstrate the applicability of this protein microarray for quantitative and simultaneous analysis of IL-1β, IL-1ra, IL-6, IL-8, MCP-1 and TNFα in clinical samples.