Niederkofler Eric E, Tubbs Kemmons A, Kiernan Urban A, Nedelkov Dobrin, Nelson Randall W
Intrinsic Bioprobes, Inc., 625 South Smith Road, Suite 22, Tempe, AZ 85281, USA.
J Lipid Res. 2003 Mar;44(3):630-9. doi: 10.1194/jlr.D200034-JLR200. Epub 2002 Dec 1.
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 microl 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 approximately 15 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.
已开发出用于分离和结构表征血浆载脂蛋白A-I（apoA-I）、载脂蛋白A-II和载脂蛋白E的新型质谱免疫分析方法（MSIA）。这些分析方法将通过亲和捕获对载脂蛋白种类进行选择性分离与使用基质辅助激光解吸/电离飞行时间质谱进行质量特异性检测相结合。在实际应用中，（从用手指采血针从个体采集的50微升全血中获取的）血浆用针对特定载脂蛋白的抗体衍生化的亲和移液器吸头进行处理。每次分析所需时间约为15分钟，如果并行对多个个体进行分析则所需时间更短。在对五名个体的简短研究中，鉴定出了几种最近报道的载脂蛋白A-II变体，并且在所有个体中均一致观察到。此外，在三名个体中观察到载脂蛋白E的E3/E3表型，在其余两名个体中观察到E2/E3和E3/E4表型，其中后者患有阿尔茨海默病。总体而言，MSIA方法提供了一种从少量血浆中分析载脂蛋白的快速、灵敏且高度准确的方法。
