Heparan sulfate phage display antibodies recognise epitopes defined by a combination of sugar sequence and cation binding

Abstract

a Phage display antibodies are widely used to follow heparan sulfate (HS) expression in tissues and cells.We demonstrate by ELISA, that cations alter phage display antibody binding profiles to HS and this is mediated by changes in polysaccharide conformation, demonstrated by circular dichroism spectroscopy. Native HS structures, expressed on the cell surfaces of neuroblastoma and fibroblast cells, also exhibited altered antibody binding profiles following exposure to low mM concentrations of these cations. Phage display antibodies recognise conformationally-defined HS epitopes, rather than sequence alone, as has been assumed, and resemble proteins in being sensitive to changes in both charge distribution and conformation following binding of cations to HS polysaccharides.

Antiplexin D1 Antibodies Relate to Small Fiber Neuropathy and Induce Neuropathic Pain in Animals

“… 6 The former was originally described in 5 IgM paraproteinemia cases presenting with a painful, predominantly sensory polyneuropathy, 26 whereas the latter was also reported in sensorimotor polyneuropathy. 27 The ubiquitous existence of these relevant antigens 28 - 30 and the absence of passive transfer experiments in animals make interpretation of their roles in SFN difficult.…”

“…the latter was also reported in sensorimotor polyneuropathy. 27 The ubiquitous existence of these relevant antigens [28][29][30] and the absence of passive transfer experiments in animals make interpretation of their roles in SFN difficult.…”

Heparan sulfate and heparin interactions with proteins

“…The effects of cation binding on conformation and dynamics have also been analysed with heparin-derived oligosaccharides possessing different substitution patterns, which exhibited considerable anisotropy and distinct internal motions. Furthermore, the recognition of HS by phage display antibodies can be altered by exposure to cations [143]. Altering the predominant cation from Na þ to Ca 2þ induced conformational changes in the 2-O-sulfated iduronic acid residue, which was transferred to the conformation of the adjacent glucosamine moiety [144].…”

Poly(ethylene glycol acrylate)‐functionalized hydrogels for heparan sulfate oligosaccharide recognition

“…Physicochemical approaches by mass or nuclear magnetic resonance spectroscopies are increasingly been used to decipher HS/heparin structures in pure samples, although these approaches are still insufficient to identify sulfation patterns or sequences on HS/heparin polysaccharidic chains . Biological approaches consider rising antibodies allowing topological recognitions of different HS/heparin domains, although the specificity of particular sequence recognition at the structural level is still far to be reached at sufficient levels . Among functional approaches, heparin—the most used anticoagulant in surgery—is controlled via anticoagulant standard tests such as thromboplastin time and activated partial thromboplastin time for proper therapeutic outcomes .…”

“…2 Biological approaches consider rising antibodies allowing topological recognitions of different HS/heparin domains, although the specificity of particular sequence recognition at the structural level is still far to be reached at sufficient levels. 3,4 Among functional approaches, heparin-the most used anticoagulant in surgery-is controlled via anticoagulant standard tests such as thromboplastin time and activated partial thromboplastin time for proper therapeutic outcomes. 5 Thus, although important advances keep ongoing, development of new approaches capable to detect specific HS/heparin fractions or sequences is still required.…”