Site-specific polymer-streptavidin bioconjugate for pH-controlled binding and triggered release of biotin

Abstract

Low molecular weight copolymers of acrylic acid (AAc) and N-isopropylacrylamide (NIPAAm) have been synthesized with reactive OH groups at one end, using a chain transfer polymerization technique. The copolymer displays both pH and temperature sensitivity over a wide and useful range of pHs and temperatures, which permits both pH and temperature control of polymer conformation. This copolymer has been conjugated to a specific cysteine thiol site inserted by genetic engineering near the recognition site of streptavidin (SAv). In this paper, we demonstrate that this bioconjugate can provide pH control of biotin binding to and triggered release from the mutant SAv. These actions are relevant to affinity separations, biosensors, diagnostics, enzyme processes, and targeted delivery of drugs or chemical agents, labels, and other signals.