Poly(vinyl alcohol-co-ethylene) (PVA-co-PE) nanofibrous membranes with surface incorporated Cibacron Blue F3GA (CB) were prepared for bio-separation and purification applications. PVA-co-PE nanofibers were fabricated via melt extrusion of an immiscible blend of PVA-co-PE polymer with cellulose acetate butyrate (CAB) as a sacrificial matrix and subsequent removal of CAB. PVA-co-PE nanofiber dispersions were coated on a releasing surface to form nanofibrous membranes with fiber sizes ranging in 50-300 nm. Cibacron Blue F3GA (CB) was covalently immobilized onto the nanofiber surfaces and reached a maximum amount of 220 mg/g on the nanofibers. The CB attached PVA-co-PE nanofibrous membranes could adsorb large amount of bovine serum albumin (BSA) (105.5 mg/g nanofiber) at pH 5.0 while the non-specific BSA adsorption on the pristine PVA-co-PE nanofibrous membrane was negligible. The elution efficiency of BSA in 1 M NaSCN was 92.5%. After 10 adsorption-elution cycles, a slight decrease of BSA capture capacity on the membrane was observed. This work demonstrated the feasibility of using the CB functionalized PVA-co-PE nanofibrous membranes in bio-separation and purification applications. (C) 2011 Elsevier B. V. All rights reserved.