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Heparan sulfate (HS) is a linear, polydisperse sulfated polysaccharidebelonging to the glycosaminoglycan family. HS proteoglycans are ubiquitouslyfound at the cell surface and extracellular matrix in animal species. HS isinvolved in the interaction with a wide variety of proteins and the regulationof many biological activities. In certain pathologic conditions, expression andshedding of HS proteoglycans is overregulated, or enzymatic degradation of HSin lysosomes is deficient, both leading to excess circulating free HS chains inblood plasma. HS has therefore been suggested as a biomarker for various severedisease states. The structural heterogeneity makes the quantification ofheparan sulfate in complex matrices such as human plasma challenging. HS plasmalevels are usually quantified by either disaccharide analysis or enzyme linkedimmunosorbent assay(ELISA). Both methods require time-consumingmultistep-protocols. We describe here the instant detection of heparan sulfatein spiked plasma samples by the Heparin Red Kit, a commercial mix-and-readfluorescence microplate assay. The method enables HS quantification in the lowmicrogram per mL range without sample pretreatment. Heparin Red appears to besufficiently sensitive for the detection of highly elevated HS levels asreported for mucopolysaccharidosis, graft versus host disease aftertransplantation, dengue infection or septic shock. This study is a significantstep toward the development of a convenient and fast method for thequantification of HS in human plasma, with the potential to simplify thedetection and advance the acceptance of HS as a biomarker.

Instant determination of the potential biomarker heparan sulfate in human plasma by a mix-and-read fluorescence assay