Article ; Online: Covalent immobilization of coagulation factor VIII on magnetic nanoparticles for aptamer development.
Journal of applied biomaterials & functional materials
2018 Volume 16, Issue 3, Page(s) 161–170
Abstract: Introduction: Magnetic nanoparticles (MNPs) are one of the most useful particulate systems in analytical applications such as specific aptamer selection. Proteins are the most noted targets of aptamer selection. Generally, covalently immobilized protein ...
Abstract | Introduction: Magnetic nanoparticles (MNPs) are one of the most useful particulate systems in analytical applications such as specific aptamer selection. Proteins are the most noted targets of aptamer selection. Generally, covalently immobilized protein coated MNPs are more stable structures. Methods: In this study, coagulation factor VIII (FVIII) was immobilized on MNPs. A silica coating provided isocyanate functional groups was considered to interact covalently with reactive groups of the protein, resulting in a stable protein immobilization. The reactions was run in dried toluene. At end, these MNPs were applied for affinity determination of a previously selected FVIII specific aptamers. Results: Immobilization of 1 mg FVIII (~ 3 nmol) on 5 mg particles was achieved with no significant particle aggregation. Using a fluorescence-based method, affinity measurement resulted in a calculated dissociation constant of 120 ± 5.6 nM for the FVIII-specific aptamer to the FVIII-coated MNPs. Conclusion: The final product could be a suitable protein-coated solid support for magnetic-based aptamer selection processes. |
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MeSH term(s) | Aptamers, Nucleotide/chemistry ; Aptamers, Nucleotide/metabolism ; Factor VIII/chemistry ; Factor VIII/metabolism ; Humans ; Immobilized Proteins/chemistry ; Immobilized Proteins/metabolism ; Isocyanates/chemistry ; Magnetite Nanoparticles/chemistry ; Microscopy, Electron, Transmission ; Protein Stability ; SELEX Aptamer Technique ; Silanes/chemistry ; Silicon Dioxide/chemistry ; Spectrometry, X-Ray Emission ; Spectroscopy, Fourier Transform Infrared ; X-Ray Diffraction |
Chemical Substances | 3-(triethoxysilyl)-propylisocyanate ; Aptamers, Nucleotide ; Immobilized Proteins ; Isocyanates ; Magnetite Nanoparticles ; Silanes ; Silicon Dioxide (7631-86-9) ; Factor VIII (9001-27-8) |
Language | English |
Publishing date | 2018-04-02 |
Publishing country | United States |
Document type | Journal Article |
ZDB-ID | 2673624-X |
ISSN | 2280-8000 ; 1722-6899 |
ISSN (online) | 2280-8000 |
ISSN | 1722-6899 |
DOI | 10.1177/2280800018765046 |
Database | MEDical Literature Analysis and Retrieval System OnLINE |
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