Applications of multifunctional magnetic nanoparticles for the enrichment of proteins for PAGE separation

Abstract

This paper describes the applications of multifunctional magnetic nanoparticles (MNPs) for the enrichment of low-abundance proteins for polyacrylamide gel electrophoresis (PAGE) separation. The hemoglobin-functionalized MNPs, named Hb-MNPs, were obtained based on electrostatic interactions and covalent binding between the hemoglobin (Hb) and the MNPs. It was demonstrated that the proteins in human serum were selectively conjugated to Hb-MNPs, which can be used for the selective enrichment of low-abundance proteins. Three and seven kinds of proteins were identified by MS after 1-D and 2-D PAGE, respectively. Comparing with native PAGE without the treatment of MNPs, some proteins were observed, such as human serum amyloid P component (SAP), vitamin D-binding protein, and serine peptidase inhibitor. Because the high concentration of SAP can be considered as a signal for the neurodegeneration of Alzheimer's disease, the present Hb-MNPs-based method was applied to investigate the serum level of SAP for the diagnosis of Alzheimer's disease, and the results are satisfying.

Enhanced Detection of Low-Abundance Human Plasma Proteins by Integrating Polyethylene Glycol Fractionation and Immunoaffinity Depletion

“…Non-denaturing electrophoresis was performed according to Sun et al [29]. The samples were re-dissolved directly in non-denaturing buffer containing 125 mM Tris-HCl, pH 6.8, 10% glycerol (v/v), and 0.001% bromophenol blue (w/v).…”

Magnetic nanoparticles for the manipulation of proteins and cells

“…Based on the principle illustrated in the work of Kriz et al, Ouyang et al made hemoglobin-functionalized MNPs that were able to enrich human serum amyloid P component (SAP), vitamin D-binding protein, and serine peptidase inhibitor. 65 Using the luminescent nanoparticles of rare earth metal oxides, Nichkova and co-workers developed a simple and robust platform for quantitative multiple protein immunoanalysis. They synthesized an interesting type of core/shell MNPs that had Co:Nd:Fe 2 O 3 as the core and luminescent Eu:Gd 2 O 3 as the shell.…”

Highly water-soluble magnetic iron oxide (Fe3O4) nanoparticles for drug delivery: enhanced in vitro therapeutic efficacy of doxorubicin and MION conjugates

“…Magnetic NPs have gained much scientic interest for their unique magnetic properties such as superparamagnetism, high coercivity, low Curie temperature, and high magnetic susceptibility. 1,2 Magnetic iron oxide nanoparticles (MIONs), owing to their advantages such as low toxicity and biocompatibility, are considered to be the most favourable candidate for bio-applications 3 including magnetic resonance imaging (MRI), 4,5 magnetic uid hyperthermia (MFH), 6 magnetic separation and immobilization of biomolecules such as nucleic acids and proteins, 7 development of drug delivery systems for controlled release of drugs, 8,9 biolabeling and magnetic sensors. 10 Thermal decomposition and co-precipitation are among the most common techniques employed for MIONs preparation owing to their advantages but these are still associated with some limitations.…”