Fabrication of Synaptotagmin II-Functionalized Neuromimetic Copolymeric Nanomembranes

Abstract

Nanoscale copolymer membranes that mimic the innate structure and properties of biological lipid membranes possessing hydrophilic and hydrophobic elements to support protein folding were used for a fundamental examination of protein—polymer integration. This study has integrated the neural synaptotagmin II (Syt II) protein, a documented target of the hemagglutinin-33 (Hn-33) protein associated with botulinum neurotoxin type A during the infection process, into polymethyloxazoline—polydimethylsiloxane—polymethyloxazoline nanomembranes. By integrating Syt II into block copolymer membranes, we have developed a neural mimetic membrane toward Hn-33 targeting the applications in nanomaterial-mediated detection. This technology can serve as a robust stand-alone platform for toxin diagnostic studies, or as a coating for integration with micro-/nanofabricated devices and electrodes for protein—protein interaction-based detection. To assess enhanced membrane complexity and toxin specificity, studies assessing the co-insertion of trisialoganglioside-GT1b (GT1b) and Syt II into the nanomembranes were used as a subsequent platform for botulinum neurotoxin type B detection. Protein—membrane integration was confirmed with atomic force microscopy imaging, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Langmuir isotherm analysis.