LIONS, NIMBE, CEA, CNRS, Université Paris-Saclay, CEA Saclay 91191, Gif sur Yvette Cedex, France.
SIMOPRO, JOLIOT, DRF, CEA, Université Paris-Saclay, CEA Saclay 91191, Gif sur Yvette Cedex, France.
Sci Rep. 2017 Jul 28;7(1):6756. doi: 10.1038/s41598-017-06660-8.
We present an automated droplet microfluidic system (DMF) to generate monitored nanoliter aqueous droplets in oil and their deposition on a commercial stainless steel plate for MALDI-TOF analysis of peptides or protein digests. We demonstrate that DMF-MALDI combination focuses the analyte on the MALDI plate, increasing considerably the homogeneity of the dried material. This results in a 30times enhanced MALDI-TOF MS signal for a model peptide, allowing a significant improvement of the detection sensitivity limit (down to few tens of attomoles). Moreover, positive detection can be achieved from sub-nanomolar peptides solutions and better overall protein sequence coverages are obtained from few tens attomoles of protein digest. These results make DMF-MALDI a promising approach for the treatment of peptides samples as well as a key component for an integrated approach in the proteomic field.
我们提出了一种自动化液滴微流控系统 (DMF)，用于在油中生成监测的纳升级水性液滴，并将其沉积在商业不锈钢板上，用于肽或蛋白质消化物的 MALDI-TOF 分析。我们证明，DMF-MALDI 组合将分析物聚焦在 MALDI 板上，极大地提高了干燥材料的均一性。这使得模型肽的 MALDI-TOF MS 信号增强了 30 倍，允许显著提高检测灵敏度极限（低至几十飞摩尔）。此外，从亚纳摩尔肽溶液中可以进行阳性检测，并且从几十飞摩尔的蛋白质消化物中可以获得更好的整体蛋白质序列覆盖率。这些结果使得 DMF-MALDI 成为处理肽样品的有前途的方法，也是蛋白质组学领域中集成方法的关键组成部分。
