Preparation of Boronic Acid-Functionalized Cryogels Using Modular and Clickable Building Blocks for Bacterial Separation

Composite cryogels containing boronic acid ligands are synthesized for effective separation and isolation of bacteria. The large and interconnected pores in cryogels enable fast binding and release of microbial cells. To control bacterial binding, an alkyne-tagged boronic acid ligand is conjugated to azide-functionalized cryogel via the Cu­(I)-catalyzed azide–alkyne cycloaddition reaction. The boronic acid-functionalized cryogel binds Gram-positive and Gram-negative bacteria through reversible boronate ester bonds, which can be controlled by pH and simple monosaccharides. To increase the capacity of affinity separation, a new approach is used to couple the alkyne-tagged phenylboronic acid to cryogel via an intermediate polymer layer that provides multiple immobilization sites. The morphology and chemical composition of the composite cryogel are characterized systematically. The capability of the composite cryogel for the separation of Gram-positive and Gram-negative bacteria is investigated. The binding capacities of the composite cryogel for Escherichia coli and Staphylococcus epidermidis are 2.15 × 10⁹ and 3.36 × 10⁹ cfu/g, respectively. The bacterial binding of the composite cryogel can be controlled by adjusting pH. The results suggest that the composite cryogel may be used as affinity medium for rapid separation and isolation of bacteria from complex samples.