The immobilization of popular chiral selectors on the surface of nanomaterials to prepare new chiral adsorbents for preparative chiral separation is a research hotspot in separation science nowadays. In this study, bovine serum albumin (BSA) modified silica nanoparticles were prepared by using polydopamine (PDA) as a versatile multifunctional secondary reaction platform. The preparation method was facile, cost-effective and environmentally friendly. The new chiral adsorbents were then investigated for the separation of representative chiral drug enantiomers. For propranolol and tryptophan, the multi-step adsorption enhanced the chiral performance to a great degree. On increasing the starting percent enantiomeric excess (%, e.e.) of the enantiomeric mixtures, the %, e.e. value of the resulting solution increased to almost 100% under the same operating conditions. For simplicity and rapidness, the results of adsorption were measured by capillary electrophoresis (CE) using carboxymethyl-β-cyclodextrin (CM-β-CD) or α-cyclodextrin (α-CD) as additives into the background electrolyte solution. The experimental results also showed that the thus-prepared nanomaterials could be readily recycled at least three times, demonstrating their great stability and possibility in practical use.