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Title: Artificial lectins : biomimetic ligands for glycoproteins
Author: Gupta, G.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2002
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This thesis describes the rational design and synthesis of selective and sterilisable biomimetic ligands for the resolution of glycosylated and carbohydrate depleted protein forms. Based on the principles of natural carbohydrate recognition, a putative library of 196 glycoprotein-binding ligands was designed and synthesised by solid phase combinatorial approaches. A preparative affinity chromatography screening assay against a well-characterised complex-type model glycoprotein, helped identify a triazine-based acyclic immobilised ligand 11/11, comprising bis-benzylamine substituents as the 'lead' ligand. The carbohydrate recognising potential of the 'lead' ligand was revealed by reduced binding of periodate oxidised model glycoprotein, and by 'sharp' elution profiles achieved with borate buffer eluents. Specific elution and competitive binding experiments of bound glycoprotein against free sugars showed that the monosaccharide specificity of the immobilised ligand was in the order mannoside>glucoside>galactoside. The ability of immobilised ligand 11/11 to bind several well-characterised oligomannose-, complex- and hybrid-type glycoproteins, and quantitatively elute them with mannoside confirmed its hexose sugar selectivity. Resolution of protein glycoforms was demonstrated by the ability of immobilised ligand 11/11 to retain and selectively elute the glycosylated form of an oligomannose-type protein, whilst the carbohydrate deficient counterpart bound non-specifically and irreversibly to the ligand. The structural integrity of immobilised ligand 11/11 was established by a semi-solution synthetic strategy, whereby the intermediate products were characterised by TLC, 1H-NMR and MS. This work has demonstrated the development of a diastereo-selective, sterilisable, stable and regulatory authority compliant biomimetic ligand, capable of resolving protein glycoforms.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available