The Purdue Make It system is a unique automated platform capable of small-scale in situ synthesis, screening small-molecule reactions, and performing direct label-free bioassays. The platform is based on desorption electrospray ionization (DESI), an ambient ionization method that allows for minimal sample workup and is capable of accelerating reactions in secondary droplets, thus conferring unique advantages compared with other high-throughput screening technologies. By combining DESI with liquid handling robotics, the system achieves throughputs of more than 1 sample/s, handling up to 6144 samples in a single run. As little as 100 fmol/spot of analyte is required to perform both initial analysis by mass spectrometry (MS) and further MSⁿ structural characterization. The data obtained are processed using custom software so that results are easily visualized as interactive heatmaps of reaction plates based on the peak intensities of m/z values of interest. In this paper, we review the system’s capabilities as described in previous publications and demonstrate its utilization in two new high-throughput campaigns: (1) the screening of 188 unique combinatorial reactions (24 reaction types, 188 unique reaction mixtures) to determine reactivity trends and (2) label-free studies of the nicotinamide N-methyltransferase enzyme directly from the bioassay buffer. The system’s versatility holds promise for several future directions, including the collection of secondary droplets containing the products from successful reaction screening measurements, the development of machine learning algorithms using data collected from compound library screening, and the adaption of a variety of relevant bioassays to high-throughput MS.

Purdue Make It 系统是一个独特的自动化平台，能够进行小规模原位合成、筛选小分子反应和进行直接无标记生物测定。该平台基于解吸电喷雾电离 (DESI)，这是一种环境电离方法，可将样品处理量降至最低，并能够加速二级液滴中的反应，因此与其他高通量筛选技术相比具有独特的优势。通过将 DESI 与液体处理机器人技术相结合，该系统实现了超过 1 个样品/秒的吞吐量，在单次运行中处理多达 6144 个样品。只需 100 fmol/点的分析物即可通过质谱 (MS) 进行初始分析和进一步的 MS ⁿ结构表征。获得的数据使用定制软件进行处理，因此基于感兴趣的m / z值的峰值强度，结果可以很容易地可视化为反应板的交互式热图。在本文中，我们回顾了之前出版物中描述的系统功能，并展示了其在两个新的高通量活动中的应用：(1) 筛选 188 种独特的组合反应（24 种反应类型，188 种独特的反应混合物）以确定反应趋势(2) 烟酰胺N的无标签研究-甲基转移酶直接来自生物测定缓冲液。该系统的多功能性为未来的几个方向提供了希望，包括从成功的反应筛选测量中收集含有产物的二次液滴，使用从化合物库筛选中收集的数据开发机器学习算法，以及将各种相关生物测定应用于高- 吞吐量 MS。