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超灵敏自制检测方法开发中的生物分析挑战:以白细胞介素-13为例的案例研究

Bioanalytical challenges in development of ultrasensitive Home Brew assays: a case study using IL-13.

作者信息

Sperinde Gizette, Hokom Martha, Peng Kun, Fischer Saloumeh K

机构信息

BioAnalytical Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.

出版信息

Bioanalysis. 2019 Jun;11(11):1045-1054. doi: 10.4155/bio-2019-0012.

Abstract

IL-13 is a biomarker of type 2 inflammation that plays a critical role in asthma. IL-13 is present in serum at subpicogram levels. Simoa HD-1 technology was evaluated for the detection and quantitation of IL-13 by using a commercially available IL-13 kit and compared with a Simoa HomeBrew (HB) IL-13 assay as well as Immunological Multi-Parameter Chip Technology (IMPACT), an internal Roche platform. Performance of the assays was evaluated based on preset criteria for sensitivity, standard curve and controls' accuracy and precision, reproducibility and parallelism of endogenous analyte in serum samples. The Simoa platform offered high assay sensitivity for evaluation of IL-13. This paper discusses the challenges and considerations when evaluating kits and/or developing HomeBrew assays using ultrasensitive platforms.

摘要

白细胞介素-13(IL-13)是2型炎症的生物标志物,在哮喘中起关键作用。IL-13以亚皮克级水平存在于血清中。使用市售的IL-13试剂盒评估了单分子阵列(Simoa)HD-1技术对IL-13的检测和定量,并与Simoa自制(HB)IL-13检测方法以及罗氏内部平台免疫多参数芯片技术(IMPACT)进行了比较。根据预设的灵敏度、标准曲线、对照的准确性和精密度、血清样本中内源性分析物的重现性和平行性标准,评估了这些检测方法的性能。Simoa平台为评估IL-13提供了高检测灵敏度。本文讨论了使用超灵敏平台评估试剂盒和/或开发自制检测方法时的挑战和注意事项。

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