cIEF of glycoproteins in short nanoporous channels

描述（由申请人提供）：迫切需要提高生物标志物发现通量的新技术。医学上有丰富的蛋白质生物标志物候选癌症，但缺乏临床验证的生物标志物。原因是发现方法的通量非常低，通常由于统计数量少而导致错误的指示。有针对性地搜索糖型是发现癌症蛋白质生物标志物的有希望的方法。我们建议确定3毫米通道填充无孔二氧化硅的可行性，用于快速等电聚焦免疫提取的血清蛋白的糖型。这些通道具有96孔格式，可无缝集成到全自动分析中，从血清到检测，每天可对数千个血清样本进行表征。之所以选择前列腺特异性抗原（PSA）作为模型蛋白，是因为有已发表的证据表明，等电聚焦可以解析糖型，具有更高的癌症特异性，而且这种生物标志物有丰富的数据和血清样本。我们已经与一位临床肿瘤学家建立了合作关系，从前列腺癌患者身上获取人类血清样本，以测试这项技术的可行性。

DESCRIPTION (provided by applicant): New technology to enable higher throughput in biomarker discovery is urgently needed. Medicine is rich in protein biomarker candidates for cancer, but poor in clinically validated biomarkers. The reason is that the methods for discovery are very low in throughput, usually giving false indications due to the statistics of small numbers Targeted searches of glycoforms are promising for the discovery of protein biomarkers for cancer. We propose to determine the feasibility of 3-mm channels packed with nonporous silica for rapid isoelectric focusing of the glycoforms of an immunoextracted protein from serum. These channels have a 96-well format for seamless integration into a fully automation analysis, from serum to detection, enabling thousands of serum samples per day to be characterized. Prostate specific antigen (PSA) is chosen as the model protein because there is published evidence that isoelectric focusing can resolve glycoforms with higher specificity for cancer, and there is a wealth of data and serum samples for this biomarker. We have established a collaboration with a clinical oncologist to obtain human serum samples from prostate cancer patients for testing the feasibility of this technology. PUBLIC HEALTH RELEVANCE: New technology will be developed to increase the discovery rate for new cancer diagnostics.