Equipe Génie Enzymatique, Membranes Biomimétiques et Assemblages Supramoléculaires, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1 - CNRS 5246 ICBMS, Bâtiment CPE - 43, bd du 11 novembre 1918, 69622 Villeurbanne, Cedex, France.
Environ Sci Process Impacts. 2013 Oct;15(10):1876-82. doi: 10.1039/c3em00296a.
The present study describes the development, optimization and performance comparison of three ELISAs and one multiplex immunoassay in a microarray format. The developed systems were dedicated to the detection of three different classes of pollutants (pesticide, explosive and toxin) in water. The characteristics and performances of these two types of assays were evaluated and compared, in order to verify that multiplex immunoassays can replace ELISA for multiple analyte sensing. 2,4-Dichlorophenoxyacetic acid, 2,4,6-trinitrotoluene and okadaic acid were chosen as model targets and were immobilized in classical microtiter plate wells or arrayed at the surface of a microarray integrated within a classical 96-well plate. Once optimized, the classical ELISAs and microarray-based ELISA performances were evaluated and compared in terms of limit of detection, IC50, linearity range and reproducibility. Classical ELISAs provided quite good sensitivity (limit of detection down to 10 μg L(-1)), but the multiplex immunoassay was proven to be more sensitive (limit of detection down to 0.01 μg L(-1)), more reproducible and an advantageous tool in terms of cost and time expenses. This multiplex tool was then used for the successful detection of the three target molecules in spiked water samples and achieved very promising recovery rates.
本研究描述了三种 ELISA 和一种微阵列格式的多重免疫分析的开发、优化和性能比较。开发的系统专门用于检测水中的三类不同污染物(农药、爆炸物和毒素)。评估和比较了这两种类型的分析的特性和性能,以验证多重免疫分析可以替代 ELISA 进行多种分析物的检测。2,4-二氯苯氧乙酸、2,4,6-三硝基甲苯和岗田酸被选为模型靶标,并固定在经典的微孔板孔中或排列在集成在经典 96 孔板中的微阵列的表面上。一旦优化,就评估和比较了经典 ELISA 和基于微阵列的 ELISA 的性能,包括检测限、IC50、线性范围和重现性。经典 ELISA 提供了相当好的灵敏度(检测限低至 10 μg L(-1)),但多重免疫分析被证明更灵敏(检测限低至 0.01 μg L(-1))、更重现性,并且在成本和时间消耗方面是一种有利的工具。然后,该多重工具用于成功检测加标水样中的三种目标分子,并获得了非常有前景的回收率。