Enzyme-Linked Cycloaddition Assay (ELCA) for rapid, ultra-sensitive monitoring of secreted sialoglycoproteins

Authors

Lundstrom, J., Yang, J., Bojar, D.

Abstract

Glycosylation of proteins is central to cell signaling, immune function, and pathogen interactions, yet existing methods for monitoring glycan changes require specialized instrumentation and primarily report on membrane-anchored, rather than secreted, glycoproteins, with a slow turnover. Here, we present the Enzyme-Linked Cycloaddition Assay (ELCA), a click chemistry-based platform for ultra-sensitive detection and semi-quantitative analysis of secreted sialoglycoproteins. By metabolically incorporating an azide-modified sialic acid into newly synthesized glycoproteins and capturing labeled material via strain-promoted cycloaddition, ELCA quantifies aggregate sialylation using a microplate reader-compatible, ELISA-like workflow. We demonstrate that the secreted glycoproteome responds rapidly to pharmacological perturbation, with changes detectable within hours. Benchmarking against common glycosylation inhibitors and profiling cytokine-driven macrophage polarization further establishes ELCA's sensitivity and temporal resolution. Compatible with serum-containing conditions and requiring no specialized instrumentation, ELCA provides a broadly accessible tool for rapid, cost-effective monitoring of secreted glycoprotein dynamics.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 20 Jun 2026.

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