Enhanced Plasma Persistence of Enzyme with Therapeutic Potential Using Polyethylene Glycol as a Coupling Agent https://doi.org/10.26538/tjnpr/v1i3.5
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Abstract
Therapeutic proteins of microbial origin though possess good pharmacological activities, gets quickly broken down into amino acids by the action of proteases in the body. Hence to protect its activity and integrity the enzyme L-arginine deiminase (ADI) was coupled with a compatible polymer methoxypolyethylene glycol-succinimidyl succinate (mPEG-SS) and was further evaluated for the presence of its activity. Enzyme production was carried out by culturing Vibrio alginolyticus under optimal conditions and was purified by standard chromatographic techniques. The purified enzyme obtained was coupled with mPEG-SS 20k (1:50 mM) and purified. Coupling
increased the molecular weight of the enzyme to about 112 kDa. The Km and Vmax values of immobilized enzyme were found to be 2.94 ± 0.13 mM and 129.87 ± 1.24 U/mL/min, respectively. Though there was a slight change in Km and Vmax values, it still retained its activity. Further, there was two-fold increase in the plasma half-lives of the enzymes. L-Arginine deiminase of Vibrio
alginolyticus remained effective even after pegylation and showed enhanced plasma persistence in normal as well as tumour bearing mice. Hence it can be a promising candidate in treatment of L-arginine auxotrophic cancers. Further studies must be carried out to develop the enzyme in the form of chemotherapeutic agent
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