The grafting of a drug molecule, i.e., geldanamycin (GA) onto polyethyleneimine (PEI)-coated magnetic nanoparticle produces a novel composite, GA@Fe3O4-NH2. The composite is confirmed by characterizations with FT-IR, Raman, SEM, EDS, VSM and TEM. Due to the high binding-affinity of GA with myosin heavy chain (MYH), GA@Fe3O4-NH2 exhibits excellent adsorption performance towards myosin. Consequently, a solid-phase extraction procedure is established for highly efficient and selective separation of myosin from pig heart extract. At pH 6.0, an adsorption efficiency of 97.1 % is achieved for treating 100 μg mL-1 myosin (0.1 mL) with 0.1 mg GA@Fe3O4-NH2 as adsorbent. The adsorption behavior of myosin onto GA@Fe3O4-NH2 fits Langmuir model, corresponding to a theoretical adsorption capacity of 518.1 mg g-1. The adsorbed myosin can be readily recycled by the SDS solution (1 %, m/m) with an elution efficiency of 91.8 %. According to circular dichroism spectroscopy, the conformational changes of myosin during adsorption and elution are reversible. For practical application, myosin is successfully isolated from the pig left ventricular protein extract with GA@Fe3O4-NH2, and SDS-PAGE and LC-MS/MS showed that myosin had high purity and a total of 716 proteins could be identified. Significantly, Geldamycin-encapsulated magnetic nanoparticle for the separation of myosin well exploits the potential of the nanomaterials modified by drug molecules in the separation and purification of target proteins.
Keywords: Geldanamycin; Magnetic nanoparticle; Myosin; Separation and purification of the proteins; Solid phase extraction.
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