High Throughput Biomarker Verification by MALDI-MS Phase II

通过 MALDI-MS 第二阶段进行高通量生物标志物验证

• 批准号: 8761855
• 负责人: RICHARD ELLSON
• 金额: $ 99.3万
• 依托单位: LABCYTE, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-25 至 2015-09-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8761855
• 关键词: Acoustics; Affinity; Biological Markers; Chemistry; Development; Lasers; Liquid substance; Malignant Neoplasms; Peptide antibodies; Peptides; Phase; Process; Proteins; Reproducibility; Sampling; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Surface; Techniques; Technology; base; biochip; liquid chromatography mass spectrometry; mass spectrometer; monolayer; multiple reaction monitoring; novel; stable isotope

Development of a novel high throughput multiplex affinity capture platform to detect low-abundance cancer related proteins from bodily fluids is proposed. The approach leverages Acoustic Droplet Ejection (ADE) technology from Labcyte Inc., recent developments in biochip fabrication, self-assembled monolayer surface chemistries, and Matrix Assisted Laser Desorption Ionization¿Mass Spectrometry (MALDI-MS) and other surface sampling mass spectrometric approaches to create a fully automated, multiplex, affinity capture platform with the reproducibility, sensitivity, and dynamic range required for routine, absolute quantitation of low-abundance cancer-related proteins. In particular, the platform will offer significantly greater throughput than Liquid Chromatography¿Mass Spectrometry (LC MS/MS) approaches such as the mass spectrometric quantitation of peptides and proteins using stable isotope standards and capture by anti-peptide antibodies (commonly called the SISCAPA process), while exploiting many of the more attractive aspects of that technique including stable isotope standards and anti-peptide antibodies. With completion of Phase II, the automated platform developed when employed in conjunction with a state-of-the-art MALDI mass spectrometer equipped with a 5 kHz laser will enable multiplex quantitation of protein biomarkers from 96 samples at a rate that is at least 10-fold faster than LC-MS/MS approaches based on Multiple Reaction Monitoring (MRM) and with comparable sensitivity, reproducibility, and dynamic range.

提出了一种新型的高通量多重亲和捕获平台，用于检测体液中低丰度的癌症相关蛋白。该方法利用Labcyte公司的声学液滴喷射（ADE）技术、生物芯片制造、自组装单层表面化学、基质辅助激光解吸电离质谱（MALDI-MS）和其他表面采样质谱方法的最新发展，创建了一个全自动、多路、亲和捕获平台，具有常规检测所需的再现性、灵敏度和动态范围。低丰度癌症相关蛋白的绝对定量。特别是，该平台将提供比液相色谱-质谱（LC MS/MS）方法更大的通量，如使用稳定同位素标准的肽和蛋白质的质谱定量和抗肽抗体捕获（通常称为SISCAPA过程），同时利用该技术的许多更有吸引力的方面，包括稳定同位素标准和抗肽抗体。随着第二阶段的完成，该自动化平台与配备5 kHz激光的最先进的MALDI质谱仪一起开发，将能够从96个样品中对蛋白质生物标志物进行多重定量，其速度至少比基于多反应监测（MRM）的LC-MS/MS方法快10倍，并且具有相当的灵敏度，再现性和动态范围。