Melanoma Proteins in Serum and Urine

血清和尿液中的黑色素瘤蛋白

• 批准号: 6737021
• 负责人: Larry W. Tjoelker
• 金额: $ 10万
• 依托单位: MACROGENICS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6737021
• 关键词: affinity labeling; biomarker; blood chemistry; diagnosis design /evaluation; early diagnosis; enzyme linked immunosorbent assay; human tissue; mass spectrometry; melanoma; neoplasm /cancer diagnosis; oncoproteins; patient oriented research; proteomics; urinalysis; western blottings

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.

描述（由申请人提供）：该提案的总体目标是在黑色素瘤患者的血清和尿液样本中鉴定可诊断癌症的特定标记蛋白或蛋白表达模式。与许多恶性肿瘤一样，早期检测对于黑色素瘤的成功治疗至关重要。质谱技术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。我们的目标是利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，提出以下具体目标： 1. 对三名黑色素瘤患者和三名正常对照的血清样本进行质谱实验。 2. 对三名黑色素瘤患者和三名正常对照的尿液样本进行质谱实验。 3. 为了发现其他生物标志物，在进行质谱分析之前，将通过其他程序（糖基化捕获和亲和基质，例如蛋白 A 和染料色谱）对样品进行分级。 4. 通过替代方法（例如，使用可用抗体或特征酶的酶活性对标记物进行蛋白质印迹或 ELISA）来确认 15 个黑色素瘤样本的生物标记物表达。 新鉴定的蛋白质的相对表达水平还将与已知的黑色素瘤标记物（例如乳酸脱氢酶）进行比较。本文描述的初步蛋白质组学实验证明，可以使用同位素编码的亲和标签（ICAT）结合串联质谱法来鉴定不同生理条件下特征的蛋白质。该提案的具体目标是将这项技术应用于识别恶性黑色素瘤的早期标志物。