Biomarker Developmental Unit

摘要 亚利桑那州立大学（ASU）生物标志物参考实验室（BRL）核心 Characterization Center（BCC）有两个主要目标。第一个是调整生物标志物中开发的测定方法， 从Discovery Laboratory（BDL）的商业测定平台到来自Meso Scale Diagnostics，LLC的商业测定平台。(MSD)为 临床验证BRL的第二个目标是参与更大的EDRN协作验证研究， 支持分析的优化和验证以及诊断方法的改进。 ASU和MSD之间的合作旨在识别和验证与肺癌相关的生物标志物 和卵巢癌血清学标志物代表了一类有前途的新型标志物，用于早期癌症检测。 这些抗体包括抗肿瘤异常蛋白的自身抗体[AAbs]、抗糖基化蛋白抗体 （AGPA），以及针对微生物的抗微生物抗体。 由于癌症和良性疾病之间的微生物差异而不同的抗原。通过推广 这些抗体是产生抗体的B细胞，这些抗体在非常低的浓度下和在一定的浓度下放大来自抗原的信号。 肿瘤发生的早期阶段，此时相应的抗原本身在肿瘤细胞中可能检测不到。 流通 亚利桑那州立大学的生物设计研究所开发了一个高通量的免疫蛋白质组学平台：核酸- 可编程蛋白质阵列（NAPPA），它取代了复杂的过程点纯化蛋白质与 一个简单的点样质粒DNA然后翻译的过程。该平台用于识别有前途的 两种癌症的候选抗体。MSD的核心专长是开发和验证强大的 多重免疫测定（直接分析物检测和血清学测定）。本提案针对 转移已建立的肺癌和卵巢癌生物标志物和BCC发现的生物标志物 BDL到MSD平台上。该平台适用于大规模验证测试和未来的临床使用。 将在MSD的生物分析实验室根据药物非临床研究质量管理规范（GLP）和ASU对样本进行检测。 每年将有多达10个生物标志物转移到BRL。BDL将进行广泛的临床研究， 验证以向下选择两种类型癌症的标记物。最后一组约10项血清学检测 和/或抗原检测试验将被验证到实验室开发试验（LDT）所需的水平， 将制定支持生产约10，000份样本的试剂盒的生产程序。 BCC BRL与EDRN成员的其他合作可能包括开发和/或扩大规模 关键试剂，如重组蛋白或单克隆抗体，抗体亲和力成熟，蛋白质 检测开发、细胞外囊泡检测、核酸检测、多重检测板开发、检测 在MSD的GLP实验室进行验证和样品测试。

Abstract The Biomarker Reference Laboratory (BRL) core of the Arizona State University (ASU) Biomarker Characterization Center (BCC) has two primary goals. The first is to adapt assays developed in the Biomarker Discovery Laboratory (BDL) to the commercial assay platform from Meso Scale Diagnostics, LLC. (MSD) for clinical validation. The second goal of the BRL is to participate in larger EDRN collaborative validation studies, with support for optimization and verification of assays and improvement of diagnostic methods. This collaboration between ASU and MSD aims to identify and validate biomarkers related to lung cancer and ovarian cancer. Serological markers represent a promising class of novel markers for early cancer detection. These include autoantibodies [AAbs] against aberrant proteins from tumors, anti-glycosylated protein antibodies (AGPA) against proteins aberrantly glycosylated in cancer, and anti-microbial antibodies against microbial antigens that differ due to microbial differences between cancer and benign disease. Through replication of antibody-producing B-cells, these antibodies amplify a signal from antigens at very low concentrations and at an early stage in tumorigenesis when the corresponding antigens may not themselves be detectable in the circulation. ASU’s Biodesign Institute developed a high-throughput immunoproteomics platform: Nucleic Acid- Programmable Protein Array (NAPPA), which replaces the complex process of spotting purified proteins with the simple process of spotting plasmid DNA followed by translation. This platform was used to identify promising antibody candidates for both cancer types. MSD’s core expertise is development and validation of robust multiplexed immunoassays (direct analyte detection and serology assays). The present proposal is directed towards transferring established lung and ovarian cancer biomarkers and biomarkers discovered by the BCC BDL onto the MSD platform. This platform is appropriate for large-scale validation testing and future clinical use. Samples will be tested in MSD’s Bioanalytical Laboratory under Good Laboratory Practice (GLP) and at ASU. Up to ten biomarkers per year will be transferred to the BRL. The BDL will perform extensive clinical validation to downselect markers for both types of cancer. Assays for the final set of approximately ten serology and/or antigen detection assays will be validated to a level required for a Laboratory Developed Test (LDT) and manufacturing procedures that would support the production of kits for ~10,000 samples will be developed. Additional collaborations of the BCC BRL with EDRN members may include development and/or scale-up of critical reagents such as recombinant proteins or monoclonal antibodies, antibody affinity maturation, protein assay development, extracellular vesicle detection, nucleic acid detection, multiplex panel development, assay validation, and sample testing in MSD’s GLP laboratory.