Melanoma Proteins in Serum and Urine

描述(由申请人提供)：这项建议的总体目标是在黑色素瘤患者的血清和尿液样本中识别特定的标记蛋白或蛋白表达模式，这些标记蛋白或蛋白表达模式可以诊断癌症。与许多恶性肿瘤一样，早期发现是成功治疗黑色素瘤的关键。质谱学的最新创新使其成为一项强大的技术，有可能彻底改变癌症诊断标记物的识别。我们的目标是使用这项技术来识别黑色素瘤患者样本中的诊断蛋白质组模式。为此，提出了以下具体目标： 1.对3例黑色素瘤患者和3例正常对照的血清样品进行了质谱学实验。 2.对3例黑色素瘤患者和3例正常对照的尿样进行了质谱学实验。 3.为了发现更多的生物标志物，样品将在质谱分析之前用额外的程序(糖基化捕获和亲和基质，如蛋白A和染料层析)进行分级。 4.用另一种方法(如免疫印迹或酶联免疫吸附试验检测抗体可用的标记物或特征酶的酶活性)，在15个黑色素瘤样本中确认生物标记物的表达。新发现的蛋白质的相对表达水平也将与已知的黑色素瘤标记物进行比较，如乳酸脱氢酶。本文描述的初步蛋白质组学实验表明，可以使用同位素编码的亲和标签(ICAT)与串联质谱仪相结合来鉴定不同生理条件下的蛋白质。这项提案的具体目的是将这项技术应用于恶性黑色素瘤的早期标记物的识别。

DESCRIPTION (provided by applicant): The overall objective of this proposal is to identify, in serum and urine samples from melanoma patients, specific marker proteins or patterns of protein expression, which are diagnostic of cancer. As with many malignancies, early detection is critical for successful treatment of melanoma. Recent innovations in mass spectrometry make it a powerful technology that has the potential to revolutionize the identification of diagnostic markers for cancer. We aim to use this technology to identify diagnostic proteomic patterns in samples from melanoma patients. To that end, the following specific aims are proposed: 1. Perform mass spectrometry experiments on serum samples from three melanoma patients and three normal controls. 2. Perform mass spectrometry experiments on urine samples from three melanoma patients and three normal controls. 3. For additional biomarker discovery, samples will be fractionated with additional procedures (glycosylation capture and affinity matrices such as Protein A and dye chromatography) prior to mass spectrometry. 4. Confirm biomarker expression with a larger panel of 15 melanoma samples by an alternative method (e.g. Western blots or ELISA for markers with available antibodies or enzymatic activity for characterized enzymes). Relative expression levels of newly identified proteins will also be compared with known melanoma markers such as lactate dehydrogenase. Preliminary proteomic experiments described herein demonstrate that proteins characteristic of different physiological conditions can be identified using isotope-coded affinity tags (ICAT) coupled with tandem mass spectrometry. The specific aims of this proposal will apply this technology to identification of early markers of malignant melanoma.