Lectin capture strategies combined with mass spectrometry for the discovery of serum glycoprotein biomarkers
Lectin capture strategies combined with mass spectrometry for the discovery of serum glycoprotein biomarkers
复制标题
DOI：
10.1074/mcp.m600176-mcp200
复制
发表时间：
2006-10-01
影响因子：
7
通讯作者：
Semmes, O. John
中科院分区：
文献类型：
--
作者：
Drake, Richard R.;Schwegler, E. Ellen;Semmes, O. John
随着技术开发、临床研究设计和生物信息学的改进，使用高通量蛋白质表达谱分析来识别血清等临床液体中的疾病生物标志物的质谱应用不断发展。以前的蛋白质表达谱研究提供了所需的洞察技术再现性，仪器校准，样品制备，研究设计和监督生物信息学数据分析的问题。在这篇综述中，新的策略，以增加临床生物标志物检测开发的蛋白质表达谱的效用进行了讨论，重点是利用基于差异凝集素的糖蛋白捕获和靶向免疫测定。不同凝集素的碳水化合物结合特异性提供了一种生物亲和方法，补充了现有的质谱仪功能，并保留了自动化的吞吐量选项。提供了使用来自前列腺癌和肝细胞癌受试者的血清样品的具体实例，沿着了将基于凝集素的方法整合到临床流体表达谱分析策略中的建议的实验策略。我们的示例工作流程结合了使用基于免疫亲和性的靶向分析方法的生物标志物发现中早期验证的必要性，该方法与上游发现技术结合良好。
The application of mass spectrometry to identify disease biomarkers in clinical fluids like serum using high throughput protein expression profiling continues to evolve as technology development, clinical study design, and bioinformatics improve. Previous protein expression profiling studies have offered needed insight into issues of technical reproducibility, instrument calibration, sample preparation, study design, and supervised bioinformatic data analysis. In this overview, new strategies to increase the utility of protein expression profiling for clinical biomarker assay development are discussed with an emphasis on utilizing differential lectin-based glycoprotein capture and targeted immunoassays. The carbohydrate binding specificities of different lectins offer a biological affinity approach that complements existing mass spectrometer capabilities and retains automated throughput options. Specific examples using serum samples from prostate cancer and hepatocellular carcinoma subjects are provided along with suggested experimental strategies for integration of lectin-based methods into clinical fluid expression profiling strategies. Our example work-flow incorporates the necessity of early validation in biomarker discovery using an immunoaffinity-based targeted analytical approach that integrates well with upstream discovery technologies.