Immobilization and characterization of novel L-asparaginase variant using functionalized multi-walled carbon nanotube

Abstract

L-asparaginase is an integral part of a multi-agent chemotherapy regimen for Acute Lymphoblastic Leukemia (ALL). The development of anti-asparaginase antibodies and safety concerns limit the use of Escherichia coli L-asparaginase (EcA), an essential component of multi-agent chemotherapy treatments for ALL. Our lab has developed L-asparaginase variants that show high activity, high stability, negligible glutaminase activity, and low immunogenicity in BALB/c mice and ALL patients. However, their half-life is still less due to degradation by proteases like Cathepsin B and Asparaginyl endopeptidase. Enzyme immobilization can decrease their degradation by proteases. Multi-walled carbon nanotubes (MWCNTs) exhibited a promising immobilization yield of 95.87% upon immobilization onto these functionalized MWCNTs (f-MWCNTs), which were activated using EDC and NHS after being functionalized with carboxyl groups. SEM, TEM, TGA, FT-IR, and Raman Spectroscopy were among the analytical methods used to validate the effective immobilization of L-asparaginase on the surface of the f-MWCNT.