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Carbodiimide-mediated immobilization of serratiopeptidase on amino-, carboxyl-functionalized magnetic nanoparticles and characterization for target delivery

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Abstract

A hybrid biomaterial of serratiopeptidase enzyme was prepared with magnetic nanoparticles (MNPs) via carboxyl and amino-functionalization and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) for direct immobilization. The average size of prepared MNPs was found to be 15.05 ± 3.06 nm. Attachment of amino and carboxyl groups was confirmed by Fourier transform infrared spectroscopy. X-ray diffraction confirmed the purity and phase integrity of Fe3O4. The MNPs and enzyme-loaded-MNPs (EMNPs) were of saturation magnetization 58 and 50 emu g−1, respectively. Thermogravimetric analysis of EDC-MNPs and EMNPs showed the presence of organic coating over MNPs. Serratiopeptidase immobilized on amino-functionalized magnetic nanoparticles showed loss of enzyme activity due to crosslinking of enzyme, while serratiopeptidase immobilized on carboxyl-functionalized magnetic nanoparticles was better and gave 115.78 mg protein g−1 MNPs, enzyme loading 168.32 U g−1 MNPs at optimized MNPs-to-enzyme ratio 1.0 mg mg−1. In vitro and in vivo studies showed that EMNPs with magnetic targeting is more effective in drug permeation and reduction in edema than free enzyme.

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Acknowledgments

Mr. Sandeep Kumar gratefully acknowledges Ministry of Human Resource Development (MHRD), Govt. of India for providing the fellowship during the study. All authors are highly thankful to Advanced Enzyme Technologies Ltd, Mumbai, for providing the enzyme serratiopeptidase. All authors are thankful for helping in the analysis of the different samples to Sophisticated Analytical Instrumentation Facility (TEM, FTIR), Punjab University, Chandigarh; Central Research Facility (SEM–EDS), Indian Institute of Technology, Ropar; Institute Instrumentation Center (AFM, VSM), Indian Institute of Technology, Roorkee. All authors are highly thankful to Prof. Samir Kumar Das, Director, National Institute of Technology (NIT), Jalandhar, for providing all grants for the study.

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  1. Department of Biotechnology, Dr. B.R. Ambedkar National Institute of Technology, G T Road Bye Pass, Jalandhar, 144011, India

    Sandeep Kumar & Asim K. Jana

  2. Department of Chemistry, Dr. B.R. Ambedkar National Institute of Technology, Jalandhar, India

    Mithu Maiti

  3. Department of Pharmaceutical Sciences, Guru Jambhshwar University of Science and Technology, Hissar, India

    Isha Dhamija

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  1. Sandeep Kumar
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Correspondence to Asim K. Jana.

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Kumar, S., Jana, A.K., Maiti, M. et al. Carbodiimide-mediated immobilization of serratiopeptidase on amino-, carboxyl-functionalized magnetic nanoparticles and characterization for target delivery. J Nanopart Res 16, 2233 (2014). https://doi.org/10.1007/s11051-013-2233-x

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