A Novel Multiplexed Assay for Rapid Antibody Screening

一种用于快速抗体筛选的新型多重检测方法

• 批准号: 8502712
• 负责人: Bradley H. Garcia
• 金额: $ 38.15万
• 依托单位: PRIMORIGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8502712
• 关键词: Address; Algorithms; Amino Acid Sequence; Amylases; Antibodies; Antigens; Beta Cell; Biological Assay; Biological Markers; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell Maturation; Cell Therapy; Cells; Cessation of life; Characteristics; Chinese Hamster Ovary Cell; Computer software; Custom; Data; Data Analyses; Data Reporting; Detection; Development; Endocrine; Endoderm; Enzyme-Linked Immunosorbent Assay; Enzymes; Factor Analysis; Funding; Generations; Genes; Glucagon; Glucokinase; Goals; Growth; Guidelines; Hormones; Hour; Human; Human Genome; Image; Immunoassay; In Vitro; Individual; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Legal patent; Letters; Linear Regressions; Marketing; Metabolism; Methods; Molecular Profiling; Mus; Nuclear; Oryctolagus cuniculus; Pancreas; Pancreatic Polypeptide; Pattern; Peptide Sequence Determination; Performance; Phase; Population; Proteins; Proteome; Rattus; Reagent; Recombinant Proteins; Reference Standards; Regenerative Medicine; Regulation; Reproduction; Research; Research Personnel; Run-On Assays; Sampling; Small Business Innovation Research Grant; Solutions; Somatostatin; Specificity; Spottings; Staging; Stem cells; Structure of beta Cell of islet; System; Testing; Time; Touch sensation; Tumor Cell Line; Universities; Validation; Variant; Wisconsin; assay development; base; cell type; cellular development; computerized data processing; cost; data acquisition; design; embryonic stem cell; extracellular; high throughput screening; induced pluripotent stem cell; internal control; islet; medical schools; miniaturize; novel; phase 1 study; phase 2 study; polyclonal antibody; progenitor; programs; prototype; public health relevance; research study; screening; software development; synthetic peptide; therapy development; tool; transcription factor; user friendly software

DESCRIPTION (provided by applicant): Primorigen Biosciences will use Phase II SBIR funds to accelerate and expand the development of new regenerative medicine cell therapies by developing a novel high throughput, low cost protein quantification system for characterizing pancreatic 2 cell and islet cell differentiation. Primorigen successfully confirmed the technical feasibility of this proposal by achieving both major goals of the Phase I project, first demonstrating (and now routinely using) its proprietary Spots on Dots" frameless microarray platform for high throughput antibody screening and characterization, and then demonstrating feasibility of developing a multiplexed protein quantitation assay for pancreatic beta cell differentiation by producing a prototype multiplexed assay with matched antibody pairs for quantifying two important markers of 2 cell differentiation (Nkx2.2 and MafB) and with 9 additional assays (Amylase 2B, FoxA2, GCK, HNF6, Isl1, NeuroD1, Neurog3, Pax4 and Pax6) in advanced stages of testing and development. In Phase II, Primorigen will complete the existing panel of assays for up to 11 markers of 2 cell differentiation (Phase I) and develop a new panel for identifying and characterizing mature islet cell populations, including 1, 2, 4, 5, and Pancreatic Polypeptide (PP) cell types. These combined tools will provide a more complete solution for tracking downstream development and maturation of pancreatic beta cells, and will be validated by obtaining independent corroboration of the performance and specificity of the assays. In addition, Primorigen will collaborate with a software developer to produce a software/flatbed scanner package that enables standardized analysis and reporting of data from assays run on the Spots on Dots" platform in a rapid, 'one touch' format.

描述(申请人提供)：Primorigen Biosciences将利用第二阶段SBIR资金，通过开发一种新的高通量、低成本的蛋白质定量系统来表征胰腺2细胞和胰岛细胞的分化，从而加速和扩大新的再生医学细胞疗法的开发。Primorigen成功地确认了这项建议的技术可行性，实现了第一阶段项目的两个主要目标，首先展示(现在经常使用)其专有斑点在Dots的无框架微阵列平台上进行高通量抗体筛选和鉴定，然后通过在测试和开发的后期阶段使用9种额外的分析方法(淀粉酶2B、FOXA2、GCK、HNF6、Isl1、Neurod1、Neurog3、Pax4和Pax6)证明了开发一种用于胰腺β细胞分化的多重蛋白质定量分析的可行性，该方法与匹配的抗体对一起用于定量两个细胞分化的两个重要标记物(Nkx2.2和MafB)。在第二阶段，Primorigen将完成现有的最多11个2细胞分化标记物的分析小组(第一阶段)，并开发一个新的小组，用于识别和表征成熟的胰岛细胞群体，包括1、2、4、5和胰腺多肽(PP)细胞类型。这些组合工具将为跟踪胰岛β细胞的下游发育和成熟提供更完整的解决方案，并将通过获得对这些分析的性能和特异性的独立证实来进行验证。此外，Primorigen将与一家软件开发商合作，生产一套软件/平板扫描仪套装，以快速、‘一触式’的格式对在Dots“平台上的斑点”上运行的化验数据进行标准化分析和报告。