Development and validation of an alpha fetoprotein immunoassay using Gyros technology

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• 作者：Allison M. Given^a ; Pamela M. Whalen^b ; Peter J. O&rsquo ; Brien^a ; Chad A. Ray^a ; ^{chad.a.ray@pfizer.com}
• 关键词：Biomarker ; Validation ; Gyros ; Alpha fetoprotein ; Immunoassay
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2012
• 期刊代码：65_07317085
• 类别：ch
• 出版时间：May-June, 2012
• 卷：64-65
• 期：Complete
• 页码：8-15
• 文件大小：1086 K

摘要

Circulating alpha fetoprotein (AFP) is a diagnostic and prognostic biomarker for hepatocellular carcinoma (HCC) with potential utility as a pharmacodynamic endpoint in rodent tumor models. This application is limited, however, by low sample volumes, highlighting the need for sensitive, sample-sparing biomarker assay methods. In order to improve the utility of AFP as an oncology biomarker, we developed a method for AFP using the Gyrolab鈩? an automated microimmunoassay platform. Commercially available antibodies were screened to identify optimal combinations that were then used in a multi-factorial design of experiments (DOE) to optimize reaction conditions. Analytical validation included assessments of accuracy and precision (A&P), and dilutional linearity/hook effect, as well as reagent and sample stability. The method is reliable, with total error, a measure of accuracy and precision, less than 30%for all concentrations tested. AFP concentrations were measurable in diseased mice and undetectable in normal mice. Therefore, this novel, low volume AFP immunoassay is suitable for pre-clinical drug development, where its miniaturized format facilitates serial sampling in rodent models of cancer.