Marko-Varga G, Ekstrom S, Heildin G, Nilsson J, Laureli T
Department of Analytical Chemistry, Lund University, Lund, Sweden.
Electrophoresis. 2001 Oct;22(18):3978-83. doi: 10.1002/1522-2683(200110)22:18<3978::AID-ELPS3978>3.0.CO;2-7.
In order to meet the expected enormous demand for mass spectrometry (MS) throughput as a result of the current efforts to completely map the human proteome, this paper presents a new concept for low-cost high-throughput protein identification by matrix assisted laser desorption/ionization-time of flight-(MALDI-TOF)-MS peptide mapping using disposable polymeric high-density nanovial MALDI target plates. By means of microfabrication technology precision engineered nanovial arrays are fabricated in polymer substrates such as polymethylmethacrylate (PMMA) and polycarbonate (PC). The target plate fabrication processes investigated were precision micromilling, cold embossing and injection moulding (work in progress). Nanovial dimensions were 300, 400 or 500 microm. Typical array densities were 165 nanovials/cm2, which corresponds to 3,300 vials on a full Applied Biosystems MALDI target plate. Obtained MALDI data displayed equal mass resolution, accuracy, signal intensity for peptide standards as compared to high-density silicon nanovial arrays previously reported by our group [7], as well as conventional stainless steel or gold targets.
为了满足当前完全绘制人类蛋白质组图谱所带来的对质谱(MS)通量的巨大预期需求,本文提出了一种通过使用一次性聚合物高密度纳米小瓶基质辅助激光解吸/电离飞行时间(MALDI-TOF)-MS肽图谱进行低成本高通量蛋白质鉴定的新概念。借助微制造技术,在诸如聚甲基丙烯酸甲酯(PMMA)和聚碳酸酯(PC)等聚合物基板中制造精密工程化的纳米小瓶阵列。所研究的靶板制造工艺包括精密微铣削、冷压印和注塑成型(正在进行的工作)。纳米小瓶尺寸为300、400或500微米。典型的阵列密度为165个纳米小瓶/平方厘米,这相当于在一块完整的应用生物系统公司的MALDI靶板上有3300个小瓶。与我们小组之前报道的高密度硅纳米小瓶阵列[7]以及传统的不锈钢或金靶相比,所获得的MALDI数据显示肽标准品的质量分辨率、准确性和信号强度相同。