DMF-MALDI: droplet based microfluidic combined to MALDI-TOF for focused peptide detection.

DMF-MALDI: droplet based microfluidic combined to MALDI-TOF for focused peptide detection.
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DOI:
10.1038/s41598-017-06660-8
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发表时间:
2017-07-28
期刊:
影响因子:
4.6
通讯作者:
Malloggi F
Malloggi F
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Mesbah K;Thai R;Bregant S;Malloggi F

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我们提出了一种自动化液滴微流控系统(DMF),以产生监测纳升的水液滴在油和它们的沉积在商业不锈钢板上的MALDI-TOF分析肽或蛋白质底物。我们证明DMF-MALDI组合将分析物聚焦在MALDI板上,大大增加了干燥材料的均匀性。这导致模型肽的MALDI-TOF MS信号增强了30倍,从而显著提高了检测灵敏度极限(低至几十阿托摩尔)。此外,可以从亚纳摩尔肽溶液中实现阳性检测,并且从几十阿托摩尔的蛋白质消化物中获得更好的总体蛋白质序列覆盖率。这些结果使DMF-MALDI成为一种有前途的肽样品处理方法,也是蛋白质组学领域综合方法的关键组成部分。
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.
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