title: Aqueous nickel-nitrilotriacetate modified Fe(3)O(4)-NH(3)(+) nanoparticles for protein purification and cell targeting.
abstract: A comprehensive totally aqueous phase synthesis of nickel-nitrilotriacetate (Ni-NTA) modified superparamagnetic Fe(3)O(4) nanoparticles is presented. The Fe(3)O(4)-NTA-Ni nanoparticles are able to perform efficient and specific purification of 6-His tagged proteins from crude cell lysates, as evidenced by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The average binding capacity, as demonstrated by streptopain (M(W) 42 kDa), is 0.23 mg/mg (protein/Fe(3)O(4)-NTA-Ni). Considering the high affinity and specificity of the binding between hexahistidine motif and Ni-NTA, Ni-NTA modified nanoparticles could act as a module to carry 6-His tagged proteins on the particle surface with molecular orientation control, since only the 6-His domain could be attached. These modularly designed functional nanoparticles enhance cancer cell targeting, as supported by the in vitro receptor mediated targeting assay using RGD-4C-6-His fusion peptide. The nanoparticles show no significant hemolysis for human blood and could be investigated further for their in vivo functional imaging applications.
authors: Institute of Oral Medicine and Molecular Medicine and Center for Micro/Nano Science and Technology, National Cheng Kung University, Tainan 701, Taiwan.
authors: Dar-Bin
authors: DB
authors: Shieh
authors: Chia-Hao
authors: CH
authors: Su
authors: Fong-Yu
authors: FY
authors: Chang
authors: Ya-Na
authors: YN
authors: Wu
authors: Wu-Chou
authors: WC
authors: Su
authors: Jih Ru
authors: JR
authors: Hwu
authors: Jyh-Horng
authors: JH
authors: Chen
authors: Chen-Sheng
authors: CS
authors: Yeh