Protein Polymer Conjugates for the Treatment of Lysosomal Storage Disorder

Farook, Isha (2022) Protein Polymer Conjugates for the Treatment of Lysosomal Storage Disorder. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Isha Farook (17MS203)) 17MS203_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (3MB)

Abstract

Gaucher disease is an autosomal recessive disorder and the most common Lysosomal storage disorder caused by the deficiency of the enzyme β-glucosidase. This enzyme hydrolyses the glucocerebroside, a type of sphingolipid, into glucose and ceramide components. The deficiency of this enzyme causes the accumulation of the substrate in the lysosome. This accumulation causes the formation of so-called Gaucher cells that are distinguished by diverting the nucleus to one side. The rest of the cell is saturated with abnormal amounts of glucosylceramide and its deacylated lysolipid, glucosylsphingosine. Although enzyme replacement therapy(ERT) is effectively applied, the instability in the blood causes the need for periodic intravenous administration that, adds to the high cost of the treatment. Polypeptides as enzyme carriers have been proposed to increase the half-life time of the systemically administered enzymes inside the body, providing protection against degradation. Thus, with the help of polyvalent M6P-based Glycopolypeptide, the enzyme can be delivered specifically to the lysosome via CI-MPR (cation-independent receptor-mediated pathway) pathway. Taking this forward, a pH-responsive acetal linker was designed, which can also act as an initiator for the ROP (ring-opening polymerization) of M6P monomer to form an azide-end functionalized M6P-Polymer. The B8CYA8 wild type of the β-Glucosidase enzyme, which has only one thiol group at its surface, is used for the thiol-ene reaction with a maleimide alkyne linker to end functionalize the β-glucosidase to contain an alkyne group. Then the protein-polymer bioconjugation using copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction was performed.

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