Conjugation of carbon coated-iron nanoparticles with biomolecules for NMR-based assay

Pavel Khramtsov; Maria Kropaneva; Il'ya Byzov; Artem Minin; Alexey Mysik; Valeria Timganova; Maria Bochkova; Mikhail Uimin; Svetlana Zamorina; Anatoly Yermakov; Mikhail Rayev

doi:10.1016/j.colsurfb.2019.01.009

Conjugation of carbon coated-iron nanoparticles with biomolecules for NMR-based assay

Colloids Surf B Biointerfaces. 2019 Apr 1:176:256-264. doi: 10.1016/j.colsurfb.2019.01.009. Epub 2019 Jan 3.

Authors

Affiliations

¹ Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia; Department of Microbiology and Immunology, Biology Faculty, Perm State National Research University, 614000, 15 Bukirev Str., Perm, Russia. Electronic address: khramtsovpavel@yandex.ru.
² Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia.
³ Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia; M.N. Mikheev Institute of Metal Physics of the Ural Branch of the Russian Academy of Sciences, 620108, 18 S. Kovalevskoy Str., Ekaterinburg, Russia.
⁴ Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia; M.N. Mikheev Institute of Metal Physics of the Ural Branch of the Russian Academy of Sciences, 620108, 18 S. Kovalevskoy Str., Ekaterinburg, Russia; Ural Federal University named after the first President of Russia B.N. Yeltsin, 620002, 19 Mira Str., Ekaterinburg, Russia.
⁵ M.N. Mikheev Institute of Metal Physics of the Ural Branch of the Russian Academy of Sciences, 620108, 18 S. Kovalevskoy Str., Ekaterinburg, Russia.
⁶ Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia; Department of Microbiology and Immunology, Biology Faculty, Perm State National Research University, 614000, 15 Bukirev Str., Perm, Russia.
⁷ Laboratory of Ecological Immunology, "Institute of Ecology and Genetics of Microorganisms of the Ural Branch of the Russian Academy of Sciences" - Branch of PSRC UB RAS, 614081, 13 Golev Str., Perm, Russia; Department of Microbiology and Immunology, Biology Faculty, Perm State National Research University, 614000, 15 Bukirev Str., Perm, Russia; Ural Federal University named after the first President of Russia B.N. Yeltsin, 620002, 19 Mira Str., Ekaterinburg, Russia.

PMID: 30623813
DOI: 10.1016/j.colsurfb.2019.01.009

Abstract

In this work, we developed and optimized conjugates of carbon-coated iron nanoparticles (Fe@C) with streptavidin and monoclonal antibodies. The conjugation procedure included two stages. First, amino groups were grafted onto the carbon shell to facilitate noncovalent sorption of bovine serum albumin (BSA). Further, the covalent attachment of proteins to the BSA layer via glutaraldehyde coupling was performed. It was established and confirmed that the synthesis procedure is reproducible and allows preparation of stable conjugates. The resulting nanoparticles are clusters of Fe@C particles coated by proteins. The size of the clusters is in the range of 100-190 nm and can be controlled via the tuning of conjugation conditions, including pH, BSA-to-Fe@C ratio, etc. Conjugates of Fe@C with streptavidin and monoclonal antibodies (sizes of approximately 140-150 nm) were synthesized. Proton T₂ relaxometry was used to detect these conjugates with very high sensitivity due to the magnetic markers, Fe@C. The relaxivity (r₂) of different conjugates varied within the range of 290-450 1/s*mM. Conjugate applicability for relaxometry-based assay was confirmed by direct detection of streptococcal protein G and biotinylated BSA in a dot immunoassay.

Keywords: Assay; Carbon-coated magnetic nanoparticles; Conjugate; Nuclear magnetic resonance.

MeSH terms

Animals
Carbon / chemistry*
Cattle
Hydrogen-Ion Concentration
Iron / chemistry*
Magnetic Resonance Spectroscopy / methods*
Nanoparticles / chemistry
Nanoparticles / ultrastructure
Particle Size
Reproducibility of Results
Serum Albumin, Bovine / chemistry*
X-Ray Diffraction

Substances

Serum Albumin, Bovine
Carbon
Iron