Application of protein lysate microarrays to molecular marker verification and quantification.
Application of protein lysate microarrays to molecular marker verification and quantification.
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DOI：
10.1186/1477-5956-3-9
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发表时间：
2005-11-10
期刊：
影响因子：
2
通讯作者：
Kapoor M
中科院分区：
文献类型：
--
作者：
Ramaswamy A;Lin E;Chen I;Mitra R;Morrisett J;Coombes K;Ju Z;Kapoor M
本研究介绍了蛋白质裂解物微阵列（LMA）技术的发展和应用，用于验证和定量的人类组织样本的蛋白质生物标志物的存在。亚皮克范围的灵敏度已经实现了LMA使用非酶蛋白检测方法。从一组质量控制实验的结果，并证明了高灵敏度和重现性的LMA方法。优化的LMA方法已被应用于验证动脉粥样硬化的疾病标志物的存在和定量。应用LMA法检测52例颈动脉内膜剥脱标本中脂蛋白[a]和载脂蛋白B100。使用相同的蛋白裂解物通过ELISA验证LMA产生的数据。通过LMA和ELISA估计的蛋白量的相关性非常显著，脂蛋白[a]的r2 ≥ 0.98（p ≤ 0.001）和载脂蛋白B100的r2 ≥ 0.94（p ≤ 0.001）。这是第一份将使用蛋白质微阵列产生的数据与ELISA（一种用于验证蛋白质生物标志物存在的标准技术）进行比较的报告。LMA技术的灵敏度、可重复性和高通量质量使其成为分析临床样品中疾病特异性蛋白标志物的潜在强大技术。
This study presents the development and application of protein lysate microarray (LMA) technology for verification of presence and quantification of human tissue samples for protein biomarkers. Sub-picogram range sensitivity has been achieved on LMA using a non-enzymatic protein detection methodology. Results from a set of quality control experiments are presented and demonstrate the high sensitivity and reproducibility of the LMA methodology. The optimized LMA methodology has been applied for verification of the presence and quantification of disease markers for atherosclerosis. LMA were used to measure lipoprotein [a] and apolipoprotein B100 in 52 carotid endarterectomy samples. The data generated by LMA were validated by ELISA using the same protein lysates. The correlations of protein amounts estimated by LMA and ELISA were highly significant, with r2 ≥ 0.98 (p ≤ 0.001) for lipoprotein [a] and with r2 ≥ 0.94 (p ≤ 0.001) for apolipoprotein B100. This is the first report to compare data generated using proteins microarrays with ELISA, a standard technology for the verification of the presence of protein biomarkers. The sensitivity, reproducibility, and high-throughput quality of LMA technology make it a potentially powerful technology for profiling disease specific protein markers in clinical samples.