Development of an automated flow chemistry affinity-based purification process for DNA-encoded chemistry

IF 2 4区化学 Q3 CHEMISTRY, MULTIDISCIPLINARY

Journal of Flow Chemistry Pub Date : 2023-09-26 DOI:10.1007/s41981-023-00282-0

Robin Dinter, Katharina Götte, Franziska Gronke, Leon Justen, Andreas Brunschweiger, Norbert Kockmann

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Abstract

An automated flow chemistry platform for DNA-encoded library (DEL) technologies requires the integration of a purification process for DNA-tagged substrates. It facilitates the development of further DEL reactions, building block rehearsal, and library synthesis. Therefore, a recently developed, manual affinity-based batch purification process for DNA-tagged substrates based on dispersive solid-phase extraction (DSPE) was transferred to automated flow chemistry using tailored 3D-printed microfluidic devices and open-source lab automation equipment. The immobilization and purification steps use Watson–Crick base pairing for a compound-encoding single-stranded DNA, which allows for the thorough removal of impurities and contaminations by washing steps and operationally simple recovery of the purified DNA-encoded compounds. This work optimized the annealing step for flow incubation and DNA purification was accomplished by flow DSPE washing/elution steps. The manually performed batch affinity-based purification process was compared with the microfluidic process by determining qualitative and quantitative DNA recovery parameters. It aimed at comparing batch and flow purification processes with regard to DNA recovery and purity to benefit from the high potential for automation, precise process control, and higher information density of the microfluidic purification process for DNA-tagged substrates. Manual operations were minimized by applying an automation strategy to demonstrate the potential for integrating the microfluidic affinity-based purification process for DNA-tagged substrates into an automated DNA-encoded flow chemistry platform.

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为 DNA 编码化学开发基于亲和力的自动流动化学纯化流程

用于 DNA 编码文库（DEL）技术的自动化流程化学平台需要整合 DNA 标记底物的纯化过程。这有助于进一步开发 DEL 反应、构件排演和文库合成。因此，利用定制的三维打印微流控装置和开源实验室自动化设备，将最近开发的基于分散固相萃取（DSPE）的DNA标记底物人工亲和批量纯化流程转移到自动化流化学中。固定和纯化步骤使用化合物编码单链 DNA 的 Watson-Crick 碱基配对，这样就可以通过洗涤步骤彻底清除杂质和污染物，并在操作上简单地回收纯化的 DNA 编码化合物。这项工作优化了流式孵育的退火步骤，并通过流式 DSPE 水洗/淘洗步骤完成 DNA 纯化。通过确定定性和定量的 DNA 回收参数，将人工批量亲和纯化过程与微流控过程进行了比较。其目的是比较批次和流动纯化过程的 DNA 回收率和纯度，以受益于微流体纯化过程在自动化、精确过程控制和更高信息密度方面的巨大潜力。通过采用自动化策略，最大限度地减少了人工操作，从而展示了将基于亲和力的DNA标记底物微流控纯化工艺整合到自动化DNA编码流动化学平台的潜力。

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来源期刊

Journal of Flow Chemistry CHEMISTRY, MULTIDISCIPLINARY-

CiteScore

6.40

自引率

3.70%

发文量

审稿时长

>12 weeks

期刊介绍： The main focus of the journal is flow chemistry in inorganic, organic, analytical and process chemistry in the academic research as well as in applied research and development in the pharmaceutical, agrochemical, fine-chemical, petro- chemical, fragrance industry.