Glycome profiling with a DNA-barcoded antibody library and microarray technology

Authors

• AZ Bhakta
• S Marglous
• KM Gillmann
• JC Gildersleeve

Abstract

Surface glycans differ between cancer and healthy cells, as well as between different types of cancer cells, but the limitations of current methods for studying surface glycosylation have led to a gap in this area of research. Methods commonly used for studying proteins cannot be applied to carbohydrates, and existing technology for studying glycans, such as mass spectrometry or lectins, often have lower throughput or specificity. Anti-glycan antibodies provide a potential alternative with their standardized structure, diverse targets, and greater potential specificity. We explore two promising applications for anti-glycan antibodies by creating a DNA-barcoded antibody library and an antibody microarray. DNA-barcoded antibodies can be used in a variety of sequencing techniques to sequence surface glycans on both a bulk and single-cell level and have the potential to be used simultaneously with RNA sequencing for a multi-omic approach to profiling cells. Preliminary sequencing of isogenic colorectal cancer cell lines SW480 and SW620 have yielded promising results, suggesting that glycan sequencing with the barcoded antibodies can identify differences between the metastatic and nonmetastatic cells’ glycomes and differentiate subpopulations within each cell line. The antibody microarray similarly allows cells, lysates, or individual molecules to be run against the library of anti-glycan antibodies to further elucidate the glycans present and identify antibodies that may be used to target them. Together, the DNA-barcoded antibody library and the antibody microarray provide a novel method of studying the glycosylation of cells, proteins, and other samples with the potential for eventual applications in cancer diagnostics.

Scientific Focus Area: Cell Biology

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