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



 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.