Molecular Biomarkers, The Biodesign Institute at Arizona State University, Tempe, AZ 85287, USA.
Proteomics Clin Appl. 2011 Aug;5(7-8):454-9. doi: 10.1002/prca.201000112. Epub 2011 Jun 8.
The purpose of the work presented herein was to develop a high-throughput assay for the quantification of human insulin in plasma samples while simultaneously detecting, with high mass accuracy, any additional variant forms of insulin that might be present in each sample.
A mass spectrometric immunoassay (MSIA) was designed in which anti-human insulin antibodies were immobilized to commercially available mass spectrometric immunoassay pipette tips and used to capture insulin and related protein variants from human plasma.
Standard curves for insulin exhibited linearity (average R(2) for three days of analysis=0.99) and assay concentration limits of detection and limits of quantification for insulin were found to be 1 and 15 pM, respectively. Estimated coefficient of variations for inter-day experiments (n=3 days) were <8%. Simultaneously, the assay was shown to detect and identify insulin metabolites and synthetic insulin analogs (e.g. Lantus). Notably, insulin variants not known to exist in plasma were detected in diabetics.
This introductory study sets a foundation toward the screening of large populations to investigate insulin isoforms, isoform frequencies, and their quantification.
本文工作的目的是开发一种高通量测定人血浆胰岛素的方法，同时利用高质量精度检测每个样本中可能存在的任何其他胰岛素变异形式。
设计了一种质谱免疫分析（MSIA），其中抗人胰岛素抗体固定在市售的质谱免疫分析吸头中，用于从人血浆中捕获胰岛素和相关蛋白变异体。
胰岛素的标准曲线呈线性（3 天分析的平均 R²=0.99），胰岛素的检测限和定量限分别为 1 和 15 pM。日内实验的估计变异系数（n=3 天）<8%。同时，该测定法还能够检测和鉴定胰岛素代谢物和合成胰岛素类似物（如来得时）。值得注意的是，在糖尿病患者中检测到了血浆中不存在的已知胰岛素变异体。
本初步研究为筛选大量人群以研究胰岛素同工型、同工型频率及其定量奠定了基础。
