Separation of peptides with polyionic nanosponges for MALDI-MS analysis.
Separation of peptides with polyionic nanosponges for MALDI-MS analysis.
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DOI：
10.1021/la802723r
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发表时间：
2009-02-03
期刊：
影响因子：
--
通讯作者：
Dyer DJ
中科院分区：
文献类型：
--
作者：
Wong VN;Fernando G;Wagner AR;Zhang J;Kinsel GR;Zauscher S;Dyer DJ
采用AIBN型自由基引发剂，以金为基底，合成了由70%聚N-异丙基丙烯酰胺（PNIPAAM）和30%聚甲基丙烯酸（PMAA）组成的聚合物刷。通过将30 pM（pH = 6.2）液滴置于聚合物刷基底上，在不到120秒内完成两种肽（缓激肽和颊癌蛋白）的分级分离。吸走含有阴离子颊癌蛋白的洗脱液用于MALDI分析，而阳离子缓激肽吸附到溶胀的阴离子刷上，随后通过向底物中加入一滴甲酸来释放。这导致刷子塌陷并释放缓激肽，很像挤压海绵;与等离子体聚合物改性的MALDI基底相比，这些纳米海绵基底表现出非常高的负载能力（>2.0 mg/ml）。椭圆偏振测量表明，互补肽吸附迅速，而那些相同的电荷不和两个馏分的MALDI-MS分析表明，这两种肽的分离。监测缓激肽随时间的吸附，并且在1分钟内85%的肽从0.5mg/ml水溶液吸附到纳米海绵上。
A polymer brush consisting of 70% poly(N-isopropylacrylamide) (PNIPAAM) and 30% polymethacrylic acid (PMAA) was synthesized from gold substrates with a grafting-from AIBN type free-radical initiator. Fractionation of two peptides, Bradykinin and Buccalin, was accomplished in less than 120 seconds by placing a 30 pM (pH∼6.2) droplet onto the polymer brush substrate. The eluant containing the anionic Buccalin is pipetted away for MALDI analysis while the cationic Bradykinin adsorbed to the swollen anionic brush and was subsequently released by adding a droplet of formic acid to the substrate. This caused the brush to collapse and release the Bradykinin, much like squeezing a sponge; these nanosponge substrates exhibited very high loading capacity (>2.0 mg/ml) compared to plasma-polymer-modified MALDI substrates. Ellipsometric measurements showed that complementary peptides adsorb rapidly while those of the same charge do not and MALDI-MS analysis of the two fractions showed separation of both peptides. The adsorption of Bradykinin was monitored over time and 85% of the peptide had been adsorbed to the nanosponge in 1 minute from a 0.5 mg/ml aqueous solution.
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