Chitosan-assisted immobilization of serratiopeptidase on magnetic nanoparticles, characterization and its target delivery

Abstract

Background: Magnetic nanoparticles (MNPs) gained attentions as universal carrier for drug delivery and for enzyme immobilization.

Purpose: Target delivery of serratiopeptidase for treatment using this enzyme and applications in drug delivery.

Method: Serratiopeptidase was immobilized on chitosan amino-functionalized MNPs by covalent bonding through glutaraldehyde. Targeting of MNPs with immobilized enzyme (EMNPs) was carried out in vitro in modified diffusion cell and in vivo in rats.

Results and discussion: MNPs and EMNPs were 15.43 ± 5.22 and 18.43 ± 3.23 nm (transmission electron microscopy), crystallite size 16.89 and 21.05 nm (X-ray diffraction) and saturation magnetization 62 and 35.2 emug⁻¹, respectively. Maximum protein and enzyme loading on EMNPs were 264 mg g⁻¹ and 325 U g⁻¹, respectively. In the molecular level, maximum 52 enzyme molecules could bind to each particle showing residual activity 68%, little effect on K_M and V_max, good storage stability. Magnetic targeting of EMNPs increased the delivery (permeation) of drug through membrane in vitro and the enhanced anti-inflammatory effect on carrageenan-induced paw oedema in rats in vivo at much lower doses of enzyme than the doses required for treatment with free enzyme.

Conclusions: The enzymatic preparation of MNPs showed enhanced effects (permeation enhancement and anti-inflammatory activity) at lower concentration with magnetic targeting.

• Carrageenan-induced paw oedema
• characterizations
• immobilization
• magnetic nanoparticles
• serratiopeptidase
• target delivery

Acknowledgements

Authors are thankful to Sophisticated Analytical Instrumentation Facility (TEM, FTIR), Punjab University, Chandigarh; Central Research Facility (XRD), Indian Institute of Technology, Ropar and Institute Instrumentation Centre (VSM), Indian Institute of Technology, Roorkee helping in analysis of different samples.