Kiernan Urban A, Addobbati Riccardo, Nedelkov Dobrin, Nelson Randall W
Intrinsic Bioprobes, Inc., 625 S. Smith Rd. Ste. 22, Tempe, Arizona 85281, USA.
J Proteome Res. 2006 Jul;5(7):1682-7. doi: 10.1021/pr0601133.
Reported in this work is the development and application of a high sensitivity mass spectrometric immunoassay for the quantitative analysis of C-reactive protein from human plasma. Multiplexed affinity retrieval devices and methodology were developed to simultaneously target retinol binding protein, C-reactive protein, serum amyloid P component, as well as an added exogenous internal reference standard (staphylococcal enterotoxin B) for subsequent MALDI-TOF MS analysis. This approach allows for semiquantitative analysis of both retinol binding protein and serum amyloid P component while performing absolute quantitative measurements of C-reactive protein. The ability to qualitatively differentiate between all three human proteins and their associated variants is also maintained. Standard curve, QC, and human plasma samples were analyzed in a high throughput manner, which performed with a CV < 15%. The resultant human plasma sample C-reactive protein quantitative measurements were then compared to those achieved with a high sensitivity latex immunoturbidimetric assay.
本研究报道了一种用于定量分析人血浆中C反应蛋白的高灵敏度质谱免疫分析方法的开发与应用。开发了多重亲和检索装置和方法，以同时靶向视黄醇结合蛋白、C反应蛋白、血清淀粉样蛋白P成分以及添加的外源性内参标准物（葡萄球菌肠毒素B），用于后续的基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）分析。这种方法允许对视黄醇结合蛋白和血清淀粉样蛋白P成分进行半定量分析，同时对C反应蛋白进行绝对定量测量。还保持了对所有三种人类蛋白质及其相关变体进行定性区分的能力。以高通量方式分析标准曲线、质量控制（QC）和人血浆样本，变异系数（CV）<15%。然后将所得人血浆样本C反应蛋白的定量测量结果与高灵敏度乳胶免疫比浊法获得的结果进行比较。
