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

 描述(申请人提供)：通过与大量的糖结合蛋白(GBP)相互作用，如C型凝集素、Galectins、Siglecs和微生物GBP，多糖在维持细胞黏附、信号转导和免疫反应等多种细胞功能中起着至关重要的作用。GBP的另一个主要类别是抗葡聚糖抗体，它是输血的关键决定因素，也可能作为包括癌症和自身免疫在内的许多疾病的诊断和预后标志物的巨大潜力。尽管Gbps至关重要，但由于缺乏强大的技术，该领域的进展一直很缓慢。固体的出现部分地缓解了这一障碍。 这极大地加深了我们对多糖功能的理解。然而，固体多糖阵列在技术上具有挑战性，不容易提供给非专家，也不是一种合适的临床工具。此外，固体多糖阵列不适合用于队列研究的大样本集的分析。因此，仍然迫切需要改进和负担得起的技术，以便同时分析大量样品和多糖。开发这样一个高吞吐量和高内容的平台将极大地加快血糖研究和将血糖发现转化为临床试验的速度。我们最近已经开始开发Luminex多路糖链阵列(LMGA)平台，该平台可以满足大规模GBP分析的所有要求。我们对48个葡聚糖进行的初步研究证明了使用LMGA同时分析多个葡聚糖(一次检测多达500个葡聚糖)的可行性，每位技术人员每天可分析1020个样本。我们还证明了LMGA具有非常高的灵敏度和特异度，成本效益高，多糖组成灵活，周转时间快，需要的样本量少。这些属性使所有生物学家都可以访问该平台，并成为临床测试的极佳选择。这款R21的主要目标是将我们的LMGA扩展到包括至少250个额外的多糖。作为积极验证过程的一部分，我们还将对来自健康对照组、1型糖尿病(T1D)和癌症患者的10,000多个可用的血清样本进行大规模的抗糖蛋白抗体研究。这些研究有望确定癌症和输血以及T1D及其并发症的发展的生物标志物候选。经过验证的LMGA将在不久的将来通过合作或服务向科学界提供各种研究。

 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.