Surfaces engineered to selectively capture and release target biomolecules and cells "on demand" are of considerable interest in a variety of biomedical applications, such as diagnostics and detection. In this work, a strategy is proposed to achieve "catch and release" using boronic acid ligand-functionalized electrospun cellulose acetate (CA) nanofiber mats. To achieve this goal, a simple method based on the one-step visible light-induced grafting copolymerization of 3-(acrylamido)phenylboronic acid (BA) and acrylamide (AM) is established. The copolymer poly(AM-co-BA)-grafted CA nanofiber mats (CA-g-P(AM-co-BA)) showed high capacity for the binding of glycoproteins such as horseradish peroxidase (HRP) and ovalbumin (OVA) under alkaline (pH 9.0) and physiological conditions (pH 7.4). Moreover, the bound glycoproteins can be released from the CA-g-P(AM-co-BA) mats by simple exposure to an acetic acid-sodium acetate (HAc-NaAc) buffer solution (pH 4.0). In addition, this system can be applied to directly extract glycoproteins from egg white samples. Likewise, the CA-g-P(AM-co-BA) mats acted as favorable substrates for Gram-negative E. coli bacterial attachment. The release of the attached bacteria is triggered by competitive sugar binding involving the addition of glucose. It can be envisioned that such boronic acid ligand-functionalized CA nanofiber mats that can "catch and release" biomolecules and bacterial cells hold considerable promise for diverse applications.