Multiplex Luminex glycan array for large scale analyses of glycan-binding proteins

 描述（由申请方提供）：聚糖通过与大量聚糖结合蛋白（GBP）（如C型凝集素、半乳糖凝集素、单细胞凝集素和微生物GBP）相互作用，在维持多种细胞功能（如细胞粘附、信号转导和免疫应答）中发挥重要作用。GBP的另一个主要类别是抗聚糖抗体，它是输血的关键决定因素，也可能具有作为许多疾病（包括癌症和自身免疫）的诊断和预后标志物的巨大潜力。尽管GBP至关重要，但由于缺乏强大的技术，该领域的进展一直缓慢。固体的出现部分地缓解了这一障碍 表面聚糖阵列，这大大提高了我们对聚糖功能的理解。然而，固体聚糖阵列在技术上具有挑战性，并且不能容易地提供给非专家，并且它不是合适的临床工具。此外，固体聚糖阵列不适用于队列研究的大样本集分析。因此，仍然迫切需要改进和负担得起的技术，用于同时分析大量样品和聚糖。这种高通量和高含量平台的开发将大大加快糖组学研究和将糖组学发现转化为临床测试。我们最近开始开发Luminex多重聚糖阵列（LMGA）平台，以满足大规模GBP分析的所有要求。我们对48种聚糖进行的试点研究证明了使用LMGA同时分析许多聚糖（单次分析多达500种聚糖）的可行性，每位技术人员每天可分析1020份样品。我们还证明了LMGA具有非常高的灵敏度和特异性，具有成本效益和聚糖组成的灵活性，具有快速的周转时间，并且需要小的样品体积。这些属性使该平台可供所有生物学家使用，并成为临床测试的绝佳选择。这个R21的主要目标是扩展我们的LMGA，以包括至少250个额外的聚糖。作为严格验证过程的一部分，我们还将对来自健康对照、1型糖尿病（T1D）和癌症患者的10，000多份可用血清样本中的抗聚糖抗体进行大规模研究。这些研究预计将确定癌症和输血的生物标志物候选者以及T1D及其并发症的发展。经过验证的LMGA将在不久的将来通过合作或服务提供给科学界进行各种研究。

 DESCRIPTION (provided by applicant): Glycans play an essential role in the maintenance of diverse cellular functions such as cell adhesion, signal transduction and immune response through interaction with a large number of glycan-binding proteins (GBPs) such as C-type lectins, galectins, siglecs and microbial GBPs. Another main category of GBPs is antiglycan antibodies that are critical determinants for blood transfusion and may also have a tremendous potential as diagnostic and prognostic markers for many diseases including cancer and autoimmunity. Despite the critical importance of GBPs, progress in this field has been slow due to the lack of powerful technologies. This obstacle was partially alleviated by advent of the solid surface glycan arrays, which greatly enhanced our understanding of glycan functions. However, solid glycan array is technically challenging and cannot be easily made available to non-experts and it is not a suitable clinical tool. Furthermore, the solid glycan arrays are not suitable for analyses of large sample sets for cohort studies. Therefore, there is still an urgent need for improved and affordable technologies for the simultaneous analyses of large numbers of samples and glycans. Development of such a high throughput and high content platform will greatly speed up glycomic studies and the translation of glycomic discoveries into clinical tests. We have recently started development of a Luminex Multiplex Glycan Array (LMGA) platform that meets all the requirements for large scale GBP analyses. Our pilot studies with 48 glycans have demonstrated the feasibility of using LMGA to simultaneously analyze many glycans (up to 500 glycans in a single assay) for 1020 samples per day per technician. We also demonstrated that LMGA has very high sensitivity and specificity, is cost-effective and flexible in glycan composition, has a rapid turn-around time, and requires small sample volume. These attributes render the platform accessible by all biologists and an excellent choice for clinical testing. The main goal of this R21 is to expand our LMGA to include at least 250 additional glycans. As part of the vigorous validation process, we will also conduct large scale studies of antiglycan antibodies in over 10,000 available serum samples from healthy controls, type 1 diabetes (T1D) and cancer patients. These studies are expected to identify biomarker candidates for cancer and blood transfusion as well as the development of T1D and its complications. Validated LMGA will be made available to the scientific community for a variety of studies via collaboration or services in the near future.