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（由申请人提供）：原始生物科学将使用II期SBIR资金来加速和扩展新的再生医学细胞疗法，通过开发一种新型的高吞吐量，低成本的低成本蛋白质定量系统来表征胰腺2细胞和胰岛细胞的表征。 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用于量化2个细胞分化的两个重要标记（NKX2.2和MAFB）的抗体对以及9个其他测定法（淀粉酶2B，FOXA2，GCK，GCK，HNF6，ISL1，NEUROD1，NEUROG1，NEUROG3，PAX4，PAX4和PAX6），在阶段II的高级阶段，阶段为2阶段的阶段，该阶段的阶段为2。 i）并开发一个新的面板，用于识别和表征成熟的胰岛细胞种群，包括1、2、4、5和胰多肽（PP）细胞类型。这些组合的工具将为跟踪胰腺β细胞的下游开发和成熟提供更完整的解决方案，并通过获得对测定的性能和特异性的独立佐证来验证。此外，Primorigen将与软件开发人员合作生产软件/平板扫描仪软件包，该软件包能够标准化分析并报告来自点上“快速平台”平台上的“快速'一触摸”格式的数据。