Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins
Novel mass spectrometric immunoassays for the rapid structural characterization of plasma apolipoproteins
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DOI：
10.1194/jlr.d200034-jlr200
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发表时间：
2003-03-01
影响因子：
6.5
通讯作者：
Nelson, RW
中科院分区：
文献类型：
--
作者：
Niederkofler, EE;Tubbs, KA;Nelson, RW
新型质谱免疫分析（MSIA）的血浆载脂蛋白A-I（apoA-I），apoA-II，和apoE的分离和结构表征已经开发。该检测方法结合了联合收割机通过亲和捕获选择性分离载脂蛋白种类与使用基质辅助激光解吸/电离飞行时间质谱法的质量特异性检测。在应用中，血浆（使用手指刺血针从个体抽取50穆尔）用亲和移液管尖端处理，所述亲和移液管尖端用针对特异性载脂蛋白的抗体衍生化。每次测定所需的时间约为15分钟，如果对多个个体进行平行测定，则所需时间更短。在一项对5名个体的简短研究中，发现了几种最近报道的apoA-II变异体，并在所有个体中观察到一致性。此外，在其中三个个体中观察到E3/E3的apoE表型，在其余两个个体中观察到E2/E3和E3/E4，后者患有阿尔茨海默病。总的来说，MSIA方法提供了一种快速、灵敏和高度准确的方法来分析小体积血浆中的载脂蛋白。
Novel mass spectrometric immunoassays (MSIAs) for the isolation and structural characterization of plasma apolipoprotein A-I (apoA-I), apoA-II, and apoE have been developed. The assays combine selective isolation of apolipoprotein species via affinity capture with mass-specific detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In application, plasma (from 50 mul of whole blood drawn from individuals, using finger lancet) was addressed with affinity pipette tips derivatized with antibodies toward the specific apolipoprotein. The time required for each assay was similar to15 min, less if assays on multiple individuals were performed in parallel. In a brief study of five individuals, several recently reported apoA-II variants were identified and observed consistently in all individuals. Additionally, the apoE phenotype of E3/E3 was observed in three of the individuals, and E2/E3 and E3/E4 observed in the remaining two individuals, the latter of whom suffers from Alzheimer's disease. Overall, the MSIA approach offers a rapid, sensitive, and highly accurate means of profiling apolipoproteins from small volumes of plasma.