Construction of copper (Ⅱ) affinity- DTPA functionalized magnetic composite for efficient adsorption and specific separation of bovine hemoglobin from bovine serum

摘要

In this work, the copper (II) affinity-DTPA (Diethylene triamine pentacetate acid) functionalized magnetic composite was prepared and as an outstanding separation platform to isolate specific bovine hemoglobin (BHb) from bovine serum. Firstly, the Fe3O4 microspheres were prepared via solvothermal method, followed by silanization with TEOS (Tetrathoxysilane) and coating with APTMS (3-Aminopropyl trimethoxysilane). Then incorporating the DTPA ligands via the amino/anhydride reaction. Finally, the target adsorbents (Fe3O4@SiO2@NH2@DTPA@Cu2+ microspheres) were obtained via charging with copper ions. The resulting materials could selectively and specifically isolate BHb with a high maximum capture capacity of 911.3 mg g(-1). The absorbed BHb was easily eluted with a recovery of 82.4% by using 0.1 M of Na2CO3-Histidine as eluent. And the real sample analysis of Fe3O4@SiO2@NH2@DTPA@Cu2+ microspheres was also tested using bovine serum, which was identified using MALDI-TOF Ms and SDS-PAGE analysis. The excellent results demonstrate that Fe3O4@SiO2@NH2@DTPA@Cu2+ microspheres would be a good bioseparation material for biomacromolecules like histidine-tagged proteins.