Determination of β-2 microglobulin levels in plasma using a high-throughput mass spectrometric immunoassay system
Determination of β-2 microglobulin levels in plasma using a high-throughput mass spectrometric immunoassay system
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DOI：
10.1021/ac010143j
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发表时间：
2001-07-15
影响因子：
7.4
通讯作者：
Nelson, RW
中科院分区：
文献类型：
--
作者：
Niederkofler, EE;Tubbs, KA;Nelson, RW
报道了一种用于直接从生物体液中分析蛋白质的高通量质谱学免疫分析系统。96孔式机器人工作站配备了抗体衍生的亲和吸管尖端，用于从样品中并行提取特定蛋白质并随后沉积到96孔阵列阵列式基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOFMS)靶上。干扰：通过选择合适的亲和管尖端衍生化化学方法将非特异性结合蛋白的影响降至最低，通过使用疏水/亲水对比靶标增强了MALDI-TOFMS的样品制备，这也提供了促进基质/分析物晶体生长的功能，自动化质谱仪用于无人值守采集数据，导致分析速度类似于100个样品/小时(生物流体-->数据)。作为说明，给出了人体血浆样品中存在的β(2)m水平的定量MSIA。
A high-throughput mass spectrometric immunoassay (MSIA) system for the analysis of proteins directly from biological fluids is reported. A 96-well-format robotic workstation equipped with antibody-derivatized affinity pipet tips was used for the parallel extraction of specific proteins from samples and subsequent deposition onto 96-well arrayed matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) targets. Interferences:from nonspecificalIy bound proteins were minimized through choice of appropriate affinity pipet tip derivatization chemistries, Sample preparation for MALDI-TOFMS was enhanced through the use of hydrophobic/hydrophilic contrasting targets, which also presented functionalities found to promote matrix/analyte crystal growth, Automated mass spectrometry was used in the unattended acquisition of data, resulting in an analysis rate of similar to 100 samples/h (biological fluid --> data). The quantitative MSIA of beta (2)m levels present in human plasma samples is given as illustration.