Maciej-Hulme, Marissa L, Leprince, Anaelle CN, Lavin, Andre, Guimond, Scott E, Turnbull, Jeremy E 
ORCID: 0000-0002-1791-754X, Pelletier, Julien, Yates, Edwin A ORCID: 0000-0001-9365-5433, Powell, Andrew K and Skidmore, Mark A
(2023)
High sensitivity (zeptomole) detection of BODIPY-labelled heparan sulfate (HS) disaccharides by ion-paired RP-HPLC and LIF detection enables analysis of HS from mosquito midguts.
ANALYTICAL METHODS, 15 (11).
pp. 1461-1469.
Abstract
The fine structure of heparan sulfate (HS), the glycosaminoglycan polysaccharide component of cell surface and extracellular matrix HS proteoglycans, coordinates the complex cell signalling processes that control homeostasis and drive development in multicellular animals. In addition, HS is involved in the infection of mammals by viruses, bacteria and parasites. The current detection limit for fluorescently labelled HS disaccharides (low femtomole; 10<sup>-15</sup> mol), has effectively hampered investigations of HS composition in small, functionally-relevant populations of cells and tissues that may illuminate the structural requirements for infection and other biochemical processes. Here, an ultra-high sensitivity method is described that utilises a combination of reverse-phase HPLC, with tetraoctylammonium bromide (TOAB) as the ion-pairing reagent and laser-induced fluorescence detection of BODIPY-FL-labelled disaccharides. The method provides an unparalleled increase in the sensitivity of detection by ∼six orders of magnitude, enabling detection in the zeptomolar range (∼10<sup>-21</sup> moles; <1000 labelled molecules). This facilitates determination of HS disaccharide compositional analysis from minute samples of selected tissues, as demonstrated by analysis of HS isolated from the midguts of <i>Anopheles gambiae</i> mosquitoes that was achieved without approaching the limit of detection.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Animals, Mammals, Culicidae, Heparitin Sulfate, Disaccharides, Chromatography, High Pressure Liquid |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 17 Apr 2023 09:46 |
| Last Modified: | 17 Apr 2023 09:46 |
| DOI: | 10.1039/d2ay01803a |
| Open Access URL: | https://doi.org/10.1039/D2AY01803A |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3169604 |
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