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Linear polyalkylamines as fingerprinting agents in capillary electrophoresis of low‐molecular‐weight heparins and glycosaminoglycans
Author(s) -
King J. Timothy,
Desai Umesh R.
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100175
Subject(s) - dermatan sulfate , capillary electrophoresis , chemistry , chondroitin sulfate , chondroitin , heparin , glycosaminoglycan , electropherogram , low molecular weight heparin , chromatography , biochemistry
Abstract Glycosaminoglycan (GAG) analysis represents a challenging frontier despite the advent of many high‐resolution technologies because of their unparalleled structural complexity. We previously developed a resolving agent‐aided capillary electrophoretic approach for fingerprinting low‐molecular‐weight heparins (LMWHs) to profile their microscopic differences and assess batch‐to‐batch variability. In this report, we study the application of this approach for fingerprinting other GAGs and analyze the basis for the fingerprints observed in CE. Although the resolving agents, linear polyalkylamines, could resolve the broad featureless electropherogram of LMWH into a large number of distinct, highly reproducible peaks, longer GAGs such as chondroitin sulfate, dermatan sulfate, and heparin responded in a highly individualistic manner. Full‐length heparin interacted with linear polyalkylamines very strongly followed by dermatan sulfate, whereas chondroitin sulfate remained essentially unaffected. Oversulfated chondroitin sulfate could be easily identified from full‐length heparin. Scatchard analysis of the binding profile of enoxaparin with three linear polyalkylamines displayed a biphasic binding profile suggesting two distinctly different types of interactions. Some LMWH chains were found to interact with linear polyalkylamines with affinities as high as 10 nM, whereas others displayed nearly 5000‐fold weaker affinities. These observations provide fundamental insight into the basis for fingerprinting of LMWHs by linear polyalkylamine‐based resolving agents, which could be utilized in the design of advanced resolving agents for compositional profiling, direct sequencing, and chemoinformatics studies.