改善 MALDI-TOF MS 配体结合分析平行化和重现性的表面预处理策略。

Surface preparation strategies for improved parallelization and reproducible MALDI-TOF MS ligand binding assays.

出版信息

ACS Appl Mater Interfaces. 2013 Jan;5(1):6-10. doi: 10.1021/am3024756. Epub 2012 Dec 24.

Abstract

Immunoassays are employed in academia and the healthcare and biotech industries for high-throughput, quantitative screens of biomolecules. We have developed monolayer-based immunoassays for MALDI-TOF MS. To improve parallelization, we adapted the workflow to photolithography-generated arrays. Our work shows Parylene-C coatings provide excellent "solvent pinning" for reagents and biofluids, enabling sensitive MS detection of immobilized components. With a unique MALDI-matrix crystallization technique we show routine interassay RSD <10% at picomolar concentrations and highlight platform compatibility for relative and label-free quantitation applications. Parylene-arrays provide high sample densities and promise screening throughputs exceeding 1000 samples/h with modern liquid-handlers and MALDI-TOF systems.

摘要

免疫分析在学术界以及医疗保健和生物技术行业中被用于高通量、定量筛选生物分子。我们已经开发了基于单层的免疫分析方法用于 MALDI-TOF MS。为了提高并行化程度，我们将工作流程改编为光刻生成的阵列。我们的工作表明，聚对二甲苯涂层可为试剂和生物流体提供出色的“溶剂固定”，从而实现对固定化成分的灵敏 MS 检测。通过独特的 MALDI 基质结晶技术，我们证明在皮摩尔浓度下常规的日内 RSD<10%，并突出了平台兼容性，适用于相对和无标记定量应用。聚对二甲苯阵列提供了高的样品密度，并有望通过现代液体处理和 MALDI-TOF 系统实现超过 1000 个样品/小时的高通量筛选。