Affinity Capture Peptides for Clinical Mass Spectrometric Assays in Plasma

用于血浆临床质谱分析的亲和捕获肽

• 批准号: 8780269
• 负责人: Shrikumar Ambujakshan Nair
• 金额: $ 28.38万
• 依托单位: AFFINERGY, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2016-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8780269
• 关键词: Affinity; Antibodies; Binding; Biological Assay; Bone Morphogenetic Proteins; Chronic Kidney Failure; Clinical; Complex; Coronary Arteriosclerosis; Degenerative polyarthritis; Detection; Development; Diabetes Mellitus; Diagnostic; Disease; Enzyme-Linked Immunosorbent Assay; Family; Family member; Fatty acid glycerol esters; Fingerprint; Fracture; Glioma; Gold; Health; Health Care Costs; Human; Immunoassay; Maintenance; Malignant Neoplasms; Marketing; Monitor; Monoclonal Antibodies; Muscle; Obesity; Pathologic Processes; Peptide Phage Display Library; Peptides; Performance; Phase; Plasma; Plasma Proteins; Play; Production; Protein Family; Proteins; Reaction; Reagent; Research; Role; Sampling; Solutions; Specificity; Spinal Fusion; Stomach Carcinoma; Testing; Therapeutic; Tissues; Transforming Growth Factors; Variant; Vascular Diseases; assay development; base; bone; bone morphogenetic protein 2; bone morphogenetic protein 4; clinically relevant; commercial application; liquid chromatography mass spectrometry; lung Carcinoma; member; myostatin; novel; overexpression; polyclonal antibody; prototype; public health relevance; research and development; standard measure; success; synthetic peptide

DESCRIPTION (provided by applicant): We will develop the first assay for multiplexed combinations of all 14 Bone Morphogenetic Proteins. Immunoassays are the "gold standard" for measuring proteins in clinical samples. However these antibody- based tests neither quantitate protein analyte variants, nor multiplex across large numbers of superfamily members. In addition, the availability of polyclonal and monoclonal antibodies is limited. We propose a novel solution to these problems, firstly by modifying a mass spectrometric immunocapture assay to use affinity capture-peptides to enrich clinically relevant protein variants. Secondly by exploitin the efficient and high- throughput production of synthetic peptides from phage display libraries as targeted capture-reagents. For proof-of-concept, we will develop a quantitative prototype assay for multiplexed combinations of all 14 Bone Morphogenetic Protein (BMP) members in human plasma. Phase II will validate multiplexed assays for the whole BMP/Transforming Growth Factor-? (TGF-?) superfamily, whose 33 members play crucial roles in development, tissue maintenance, and many diseases. TGF-? superfamily members relevant to diagnostic and therapeutic applications include Myostatin (inhibition increases muscle and bone and decreases fat), BMP-2 (overexpressed in lung carcinomas, gastric carcinomas, and gliomas), BMP-4 (elevated in chronic kidney disease and coronary artery disease), BMPs 2 and 4 (overexpressed in degenerative joint disease), TGFs-? 1, 2 and 3 (complex involvement in cancer), and BMPs 2 and 7 (approved for nonunion bone fractures and spinal fusions). These new multiplexed assays will enable reliable, specific and faster detection of single or selected combinations of members of this large and important family, for the R&D, diagnostics and therapeutics markets.

描述（由申请方提供）：我们将开发所有14种骨形态发生蛋白多重组合的第一种检测方法。免疫分析是测量临床样品中蛋白质的“金标准”。然而，这些基于抗体的测试既不能定量蛋白质分析物变体，也不能跨大量超家族成员进行多重检测。此外，多克隆和单克隆抗体的可用性是有限的。我们提出了一种新的解决方案，这些问题，首先通过修改质谱免疫捕获测定使用亲和捕获肽，以丰富临床相关的蛋白质变体。第二，利用噬菌体展示文库高通量生产合成肽作为靶向捕获试剂。为了验证概念，我们将开发一种用于人血浆中所有14种骨形态发生蛋白（BMP）成员多重组合的定量原型测定。第二阶段将验证整个BMP/转化生长因子-？(TGF-?)超家族，其33个成员在发育、组织维持和许多疾病中起着至关重要的作用。TGF-？与诊断和治疗应用相关的超家族成员包括肌生长抑制素（抑制增加肌肉和骨并减少脂肪）、BMP-2（在肺癌、胃癌和神经胶质瘤中过表达）、BMP-4（在慢性肾病和冠状动脉疾病中升高）、BMP 2和4（在退行性关节疾病中过表达）、TGF-？1、2和3（复杂的癌症参与），以及BMP 2和7（批准用于骨折不愈合和脊柱融合）。这些新的多重检测将能够可靠、特异和更快地检测这个重要的大家族的单一或选定的组合成员，用于研发、诊断和治疗市场。