Functional characterization of recombinant human granulocyte colony stimulating factor (hGMCSF) immobilized onto silica nanoparticles.

Publisher: Kluwer Academic Publishers Country of Publication: Netherlands NLM ID: 8008051 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1573-6776 (Electronic) Linking ISSN: 01415492 NLM ISO Abbreviation: Biotechnol Lett Subsets: MEDLINE

Publication: 1999- : Dordrecht : Kluwer Academic Publishers
Original Publication: [Kew, Eng., Science and Technology Letters]

Drug Carriers* ; Nanoparticles* ; Silicon Dioxide*; Granulocyte Colony-Stimulating Factor/*metabolism ; Immobilized Proteins/*metabolism; Adsorption ; Animals ; Cell Proliferation/drug effects ; Circular Dichroism ; Cloning, Molecular ; Dynamic Light Scattering ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Gene Expression ; Granulocyte Colony-Stimulating Factor/chemistry ; Granulocyte Colony-Stimulating Factor/isolation & purification ; Humans ; Immobilized Proteins/chemistry ; Immobilized Proteins/isolation & purification ; Macrophages/drug effects ; Macrophages/physiology ; Mice ; Molecular Weight ; RAW 264.7 Cells ; Recombinant Proteins/chemistry ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/metabolism ; Spectrophotometry, Infrared

Objectives: Granulocyte macrophage colony stimulating factor (GMCSF), an important therapeutic cytokine, was immobilized onto silica nanoparticles. Maintenance of structural integrity and biological performance in immobilized cytokine was assessed to augment its applicability in possible biomedical implications.
Results: Following its cloning and expression in E. coli, the recombinant human GMCSF (hGMCSF) was purified as a GST-tagged protein corresponding to a 42 kDa band on SDS-PAGE. The purified cytokine was immobilized onto biocompatible silica nanoparticles (~129.4 nm) by adsorption and the binding was confirmed by dynamic light scattering and infrared spectroscopy. Maximum binding of hGMCSF was at 6.4 µg mg(-1) silica nanoparticles. Efficient release of the cytokine from the nanoparticles with its structural integrity intact was deduced from circular dichroism spectroscopy. hGMCSF-immobilized silica nanoparticles efficiently increased the proliferation of RAW 264.7 macrophage cells with 50 % increase in proliferation at 600 ng hGMCSF µg(-1) silica nanoparticles.
Conclusions: Silica nanoparticles successfully immobilized hGMCSF maintaining its structural integrity. The release of the immobilized cytokine from silica nanoparticles resulted in the increased proliferation of macrophages indicating the potential of the system in future applications.

Keywords: Immobilization; Macrophage proliferation; Recombinant hGMCSF; Silica nanoparticles; Therapeutic cytokine

0 (Drug Carriers)
0 (Immobilized Proteins)
0 (Recombinant Proteins)
143011-72-7 (Granulocyte Colony-Stimulating Factor)
7631-86-9 (Silicon Dioxide)

Date Created: 20151110 Date Completed: 20161101 Latest Revision: 20161230

20250114

10.1007/s10529-015-1984-0

26549378