affinity capture and detection of immunoglobulin E in human serum using an aptamer-modified surface in matrix-assisted laser desorption/ionization mass spectrometry
affinity capture and detection of immunoglobulin E in human serum using an aptamer-modified surface in matrix-assisted laser desorption/ionization mass spectrometry
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DOI：
10.1021/ac061256b
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发表时间：
2007-01-01
影响因子：
7.4
通讯作者：
McGown, Linda B.
中科院分区：
文献类型：
--
作者：
Cole, Jacquelyn R.;Dick, Lawrence W., Jr.;McGown, Linda B.
捕获和检测免疫球蛋白E（IgE）在简单的解决方案和人血清中使用适配体修饰的探针表面的亲和基质辅助激光解吸/电离质谱检测报告。在单次1 μ L应用1 pM IgE或在10次连续1 μ L应用100 fM IgE后，应用1 μ L IgE获得可检测信号。在这两种情况下，在每次应用IgE后冲洗表面以去除样品伴随物，包括盐和游离或非特异性结合的蛋白质。证明了人血清中天然IgE（血清免疫球蛋白中丰度最低且发生在亚纳摩尔水平）的检测，并研究了高丰度免疫球蛋白和白蛋白的干扰。适体修饰的表面显示出对血清中免疫球蛋白的高选择性，没有明显的血清白蛋白干扰。向血清中加入IgE抑制了来自其他免疫球蛋白的信号，证实了适体表面对IgE的预期选择性。血清稀释增加了对IgE的选择性;在10 000倍稀释的血清中检测到蛋白质而无干扰，这与标准品溶液中阿莫尔（pM）的检测一致。
Capture and detection of immunoglobulin E (IgE) in simple solution and in human serum using an aptamer-modified probe surface for affinity matrix-assisted laser desorption/ionization mass spectroscopy detection is reported. Detectable signals were obtained for 1 amol of IgE applied either in a single, 1-mu L application of 1 pM IgE or after 10 successive, 1-mu L applications of 100 fM IgE. In both cases, the surface was rinsed after each application of IgE to remove sample concomitants including salts and free or nonspecifically associated proteins. Detection of native IgE, which is the least abundant of the serum immunoglobulins and occurs at subnanomolar levels, in human serum was demonstrated and interference from the high-abundance immunoglobulins and albumin was investigated. The aptamer-modified surface showed high selectivity toward immunoglobulins in serum, with no significant interference from serum albumin. Addition of IgE to the serum suppressed the signals from the other immunoglobulins, confirming the expected selectivity of the aptamer surface toward IgE. Dilution of the serum increased the selectivity toward IgE; the protein was detected without interference in a 10 000-fold dilution of the serum, which is consistent with detection of IgE at amol (pM) levels in standard solutions.