Glycan Proximity Sequencing (GPS) For Single-Cell Multi-omics Glycoproteomics

SUMMARY

TL;DR: Glycan Proximity Sequencing (GPS) is a next-generation single-cell multi-omics assay that measures mRNA expression, extracellular protein abundance, global glycan motifs, and protein-specific glycoforms on individual cells. By combining lectin– and antibody–based proximity ligation with standard single-cell RNA sequencing workflows, GPS creates DNA barcodes that report not only which proteins and glycans are present, but which glycans decorate which proteins on each cell.

The Unmet Need: A scalable, high-resolution platform capable of measuring gene expression, proteins, and decoding protein-specific glycosylation at the single-cell level

• Single-cell multi-omics technologies have revolutionized our understanding of cellular heterogeneity by enabling the simultaneous measurement of various molecular modalities, such as gene expression and protein abundance, within individual cells. One critical aspect of cellular biology that remains challenging to profile at single-cell resolution is protein glycosylation—a post-translational modification that profoundly influences protein function, cell signaling, and immune recognition. Glycosylation patterns are highly dynamic and cell-type-specific, playing pivotal roles in processes such as immune response, cancer progression, and therapeutic efficacy. The ability to comprehensively map glycosylation alongside transcriptomic and proteomic data in single cells is essential for unraveling the complex regulatory networks that govern cell behavior and disease mechanisms.

• Despite the importance of glycosylation, current approaches to glycoproteomics face significant limitations. Traditional mass spectrometry-based methods, while powerful for bulk analysis, are inherently low-throughput, require large amounts of starting material, and lack the sensitivity to detect low-abundance glycoforms or resolve heterogeneity at the single-cell level. These approaches also struggle to directly link glycan modifications to specific proteins and their corresponding gene expression profiles within individual cells. Affinity-based assays, such as flow cytometry with lectin staining, provide some information on glycan abundance but are limited in multiplexing capacity and cannot resolve protein-specific glycoforms or integrate with transcriptomic data. As a result, there is a critical need for new technologies that can overcome these barriers and provide integrated, high-resolution, and scalable profiling of glycosylation and other molecular modalities in single cells.

The Proposed Solution: Glycan Proximity Sequencing (GPS) to unify transcriptome, proteome, glycome, and glycoproteome quantification in individual cells

• The faculty inventor developed Glycan Proximity Sequencing (GPS), an innovative single-cell multi-omics platform that enables the simultaneous measurement of mRNA expression, extracellular protein abundance, global glycan profiles, and protein-specific glycoforms within individual live cells. This technology integrates proximity ligation assay (PLA) chemistry with high-throughput single-cell RNA sequencing. It utilizes DNA-tagged antibodies and lectins as affinity probes, which, when bound in close proximity on the same glycoprotein, allow their DNA tags to be ligated into a unique fragment. This ligated DNA, along with cellular mRNA, is captured during single-cell isolation and quantified using next-generation sequencing. GPS thus provides comprehensive, parallel profiling of transcriptomic, proteomic, glycomic, and glycoproteomic modalities at single-cell resolution, with scalability determined by the number of antibody and lectin probes used.

• What sets GPS apart is its ability to overcome the limitations of traditional mass spectrometry-based glycoproteomics, which are typically low-throughput, insensitive to low-abundance glycoforms, and require bulk cell material. GPS achieves high sensitivity and throughput, enabling the detection of cell-type- and condition-specific glycosylation patterns that are critical for understanding immune function, disease progression, and therapeutic responses. Unlike previous proximity sequencing methods, GPS uniquely links protein glycosylation states to gene expression and protein abundance within the same cell, revealing dynamic remodeling events and functional associations that were previously inaccessible. Its versatility extends to profiling other post-translational modifications and integrating with spatial transcriptomics, making GPS a powerful tool for biomarker discovery, immune monitoring, and translational research in health and disease.

ADVANTAGES

ADVANTAGES

• Multi-omics integration in a single assay
• Single-cell and high-throughput compatible
• Sensitive and quantitative with strong concordance to flow cytometry and MS
• Integrates transcriptomic and glycoproteomic data to reveal cell-type- and condition-specific glycosylation patterns and their functional implications
• Scalable multiplexing of protein-glycan pairs
• Flexible platform with expansion paths

APPLICATIONS

• Single-cell glycoproteomics reagent kits and services
• Liquid biopsy and minimally invasive diagnostics
• Immuno-oncology biomarker discovery and patient stratification
• Cell therapy characterization and quality control
• Autoimmune and inflammatory disease research tools
• Expansion to multi-PTM systems biology