Microfluidic chips for protein differential expression profiling
Microfluidic chips for protein differential expression profiling
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DOI：
10.1002/elps.200800653
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发表时间：
2009-04-01
期刊：
影响因子：
2.9
通讯作者：
Lazar, Iulia M.
中科院分区：
文献类型：
--
作者：
Armenta, Jenny M.;Dawoud, Abdulilah A.;Lazar, Iulia M.
利用新的蛋白质组学技术发现和筛选生物标志物是生物医学界日益关注的一个领域。早期发现异常的生理状况将非常有利于诊断各种疾病和提高生存率。显然，这一领域的进展将取决于快速、可靠、高灵敏度和特异性样品生物分析方法的发展。微流体技术已经成为蛋白质组学研究中必不可少的技术，因为它能够整合所有样品制备，分离和检测步骤，并具有增强样品通量的额外好处。这些优点与MS检测的灵敏度和提供精确结构信息的能力相结合，使微流体-MS成为生物标志物发现的非常有竞争力的技术。本文报道了LC微芯片装置与MS检测的集成，特别是它们在MCF-7乳腺癌细胞提取物中生物标志物筛选应用的适用性。在芯片上加载类似于0.1-1 μ g粗蛋白提取物胰蛋白酶消化物通常导致可靠地鉴定类似于40-100种蛋白质。首次探索了LC-ESI-MS芯片用于同位素标记的MCF-7乳腺癌细胞提取物的比较蛋白质组学分析的潜力。
Biomarker discovery and screening using novel proteomic technologies is an area that is attracting increased attention in the biomedical community. Early detection of abnormal physiological conditions will be highly beneficial for diagnosing various diseases and increasing survivability rates. Clearly, progress in this area will depend on the development of fast, reliable, and highly sensitive and specific sample bioanalysis methods. Microfluidics has emerged as a technology that could become essential in proteomics research as it enables the integration of all sample preparation, separation, and detection steps, with the added benefit of enhanced sample throughput. The combination of these advantages with the sensitivity and capability of MS detection to deliver precise structural information makes microfluidics-MS a very competitive technology for biomarker discovery. The integration of LC microchip devices with MS detection, and specifically their applicability to biomarker screening applications in MCF-7 breast cancer cellular extracts is reported in this manuscript. Loading similar to 0.1-1 mu g of crude protein extract tryptic digest on the chip has typically resulted in the reliable identification of similar to 40-100 proteins. The potential of an LC-ESI-MS chip for comparative proteomic analysis of isotopically labeled MCF-7 breast cancer cell extracts is explored for the first time.