Quantifying proteins in plasma to democratize personalized medicine for patients with type 1 diabetes

量化血浆中的蛋白质以使 1 型糖尿病患者的个性化医疗民主化

• 批准号: 10396811
• 负责人: ANDREW N HOOFNAGLE
• 金额: $ 27.49万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396811
• 关键词: Acute; Affect; Affinity; Algorithms; Amputation; Antibodies; Antibody Specificity; Autoimmune Diseases; Award; Biological Assay; Biological Markers; Blindness; Blood; Blood Glucose; Blood Tests; C-Peptide; COVID-19 pandemic; Clinical; Clinical Trials; Companions; Data; Detection; Development; Diabetes Mellitus; Disease; Emotional; Enteroglucagon; Equilibrium; Event; Financial Hardship; Flowers; Frustration; Funding; Future; Glucagon; Glucose; Glycosylated hemoglobin A; Goals; Grant; Hybridomas; Hyperglycemia; Hypoglycemia; Immunoassay; Incidence; Injectable; Institution; Insulin; Insulin-Dependent Diabetes Mellitus; Iowa; Kidney Diseases; Laboratories; Lead; Life; Liquid Chromatography; Long-Term Effects; Manufacturer Name; Mass Spectrum Analysis; Measurement; Methods; Myocardial Infarction; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Outcome; Pancreas; Patient Care; Patients; Peptides; Periodicity; Plasma; Procedures; Process; Proinsulin; Proteins; Proteomics; Reagent; Regimen; Reproducibility; Research; Research Personnel; Risk; Signal Transduction; Somatostatin; Specificity; Structure of beta Cell of islet; Technology; Technology Transfer; Time; Translating; Translations; United States; Validation; Work; analog; base; burden of illness; cardiovascular disorder risk; clinical care; cohort; cost; effective therapy; experimental study; improved; interest; method development; novel; operation; personalized medicine; prevent; proglucagon; response; standard of care; stem; tandem mass spectrometry; technology validation; web site; website development

ABSTRACT Type 1 diabetes affects more than 1.25 million people in the United States and the annual incidence is increasing at an alarming rate of 3-4%. The emotional and financial burden of the disease is overwhelming and we currently have no way to predict or prevent new cases. As we gain a better understanding of the pathophysiological processes in the pancreas and the downstream effects of hyperglycemia (and periodic hypoglycemia during treatment), more robust biomarker assays are needed to improve the reproducibility of research findings and to translate those findings to clinical care. One technology that can provide robust, transferable assays for the measurement of proteins is liquid chromatography-tandem mass spectrometry. By directly detecting the analyte of interest, assays that use mass spectrometry detection can have better specificity than immunoassays and when paired with enrichment strategies, they can also be very sensitive. As we have demonstrated previously, it is straightforward to harmonize the results of mass spectrometric assays, which is significantly more difficult for immunoassays in general. This proposal aims to generate and validate novel transferable protein assays that harness the power of mass spectrometry. Whenever possible, assays will be multiplexed and if affinity reagents are required, they will be widely distributed through the Iowa Hybridoma Bank. Chromatographic data from method development (particularly peptide selection, which will use narrow-window data-independent acquisition rather than relying on algorithms or data-dependent acquisition methods) as well as chromatographic data from method validation will be distributed via Panorama, Chorus, and Passport, as will detailed standard operating procedures. As requested in RFA DK-17-019, five of the assays produced will target c-peptide, insulin, glucagon, glycated soluble CD59, and somatostatin/prosomatostain. Our Target Prioritization Committee will help identify the most important proteins to add to this list and focus our development efforts.

抽象的 1型糖尿病在美国影响超过125万人和年度 发病率以3-4％的惊人速度增加。情感和经济负担 疾病是压倒性的，我们目前无法预测或预防新病例。像我们 更好地了解胰腺和胰腺的病理生理过程 高血糖（治疗期间周期性低血糖）的下游影响，更多 需要强大的生物标志物分析以提高研究发现的可重复性和 将这些发现转化为临床护理。一项可以提供强大，可转移的技术 测量蛋白质的测定是液相色谱串联质谱法。 通过直接检测感兴趣的分析物，使用质谱检测的测定可以 具有比免疫测定更好的特异性，并且与富集策略配对时， 也可能非常敏感。正如我们之前所展示的那样，它很简单 协调质谱测定的结果，这对于更难 一般而言，免疫测定。该建议旨在生成和验证可转移的新颖 蛋白质测定法，利用质谱的力量。只要可能，测定将是 多路复用，如果需要亲和力试剂，它们将通过爱荷华州广泛分布 杂交瘤银行。方法开发的色谱数据（尤其是肽 选择，它将使用窄窗户依赖数据而不是依靠 算法或数据依赖性采集方法）以及色谱数据 方法验证将通过全景，合唱和护照分发，如 标准操作程序。按照RFA DK-17-019的要求，其中五个测定 将靶向C肽，胰岛素，胰高血糖素，可溶性CD59和 生长抑素/prosomatostain。我们的目标优先委员会将有助于确定最多的 重要的蛋白质可以添加到此列表中并集中我们的开发工作。

项目成果

Multiplexed quantification of insulin and C-peptide by LC-MS/MS without the use of antibodies.

• DOI: 10.1016/j.jmsacl.2022.06.003
• 发表时间: 2022-08
• 期刊: JOURNAL OF MASS SPECTROMETRY AND ADVANCES IN THE CLINICAL LAB
• 影响因子: 2.2
• 作者: Foulon, North;Goonatilleke, Elisha;MacCoss, Michael J.;Emrick, Michelle A.;Hoofnagle, Andrew N.
• 通讯作者: Hoofnagle, Andrew N.