A Thiol-functionalized silica hollow nanospheres(denoted as SiO2-SH NSs) were prepared through a hydrothermal route. The SiO2-SH NSs were conjugated with glutathione group (denoted as-GSH) to afford SiO2-GSH NSs. The as-prepared SiO2-GSH sample has hollow structure and exhibits an average diameter of about 45 nm and a wall thickness of 10 nm. The SiO2-GSH NSs were used to separate three kinds of GST-tagged proteins (GST-tagged GPX, GST-tagged LOV and GST-tagged 210-6P). These SiO2-GSH NSs exhibit negligible non-specific adsorption, high binding capacity (89.9 mu mol/g), the low detection limit (1.0 x 10(-6) mol/L) and reuse property, showing great potentiality in purifying GST-tagged proteins. (C) 2016 Elsevier Ltd. All rights reserved.