Ultra-Fast Glyco-Coating of Non-Biological Surfaces

Abstract

The ability to glycosylate surfaces has medical and diagnostic applications, but there is no technology currently recognized as being able to coat any surface without the need for prior chemical modification of the surface. Recently, a family of constructs called function-spacer-lipids (FSL) has been used to glycosylate cells. Because it is known that lipid-based material can adsorb onto surfaces, we explored the potential and performance of cell-labelling FSL constructs to “glycosylate” non-biological surfaces. Using blood group A antigen as an indicator, the performance of a several variations of FSL constructs to modify a large variety of non-biological surfaces was evaluated. It was found the FSL constructs when optimised could in a few seconds glycosylate almost any non-biological surface including metals, glass, plastics, rubbers and other polymers. Although the FSL glycan coating was non-covalent, and therefore temporary, it was sufficiently robust with appropriate selection of spacer and surface that it could capture anti-glycan antibodies, immobilize cells (via antibody), and withstand incubation in serum and extensive buffer washing, making it suitable for diagnostic and research applications.

Biofunctionalizing nanofibers with carbohydrate blood group antigens

“…Although these features are not important for achieving the aims of this article, they were incorporated critical design factors to allow use of these constructs on biological surfaces and in therapeutic applications . Potential limitations of these constructs to modify surfaces including plastics, metals are discussed elsewhere . Introduction of the desired properties of water dispersibility and self assemble required the development of a series of different spacers, in particular so called CMG (carboxymethylglycine containing spacer, ‐[NHCH 2 CO‐NHCH 2 CO‐N(CH 2 COOH)CH 2 CO‐NHCH 2 CO‐NHCH 2 CO‐N(CH 2 COOH)CH 2 CO‐NHCH 2 ‐] 2 Scheme ).…”

“…Here, we report on the ability of a new technology, Kode™ Technology, to rapidly modify electrospun nanofiber membranes with glycans. Kode™ Technology is normally used for coating cells and viruses with bioactive compounds and utilizes water dispersible self‐assembling amphiphatic function‐spacer‐lipid constructs to modify almost any surfaces with a water resistant biofunctional coating . Although only glycans were used in this initial study, as these have the potential to induce biological activity to surfaces, the potential also exists to use other function‐spacer‐lipid constructs bearing other non‐glycan functional moieties for bioactivity …”

Applications for kodecytes in immunohaematology

“…Equally, the attachment of the Kode constructs to hydrophobic surfaces can be explained in a similar manner with the hydrophobic lipid tail directly interacting with the hydrophobic surface . More complex is the ability of Kode constructs to label hydrophilic surfaces . This spontaneous self‐assembly on to almost any hydrophilic surface is believed to driven by both the hydrophobic lipid tail and the hydrophilic spacer working together to cause the amphipathic Kode construct to tightly self‐assemble on the surface.…”

“…The mechanism of surface attachment and retention is complex, surface‐type dependent and probably involves a combination of multiple forces including van der Waals, electrostatic, hydration and steric . Once attachment has occurred, the primary Kode coating is relatively water resistant . From a methodological perspective, the difference between labelling a hydrophobic surface and a hydrophilic surface is not readily observable, and both occur within seconds .…”

Rapid one-step biotinylation of biological and non-biological surfaces

“…Recently a series of water-dispersible amphipathic constructs known as function-spacer-lipids (FSLs) have been described for labelling cells, membrane viruses, and solid surfaces with a range of biological and non-biological markers under physiological conditions 3 , 4 . The basic structure of the FSL construct is typically a hydrophilic functional head; a biologically inert hydrophilic spacer which assists the construct to disperse in water and spaces the functional head away from the surface; and a lipid tail which makes the construct amphipathic and drives the spontaneous self-assembly on surfaces and insertion into lipid-based membranes 3 .…”