1. Immobilization of Proteins with Controlled Load and Orientation.
- Author
-
Bednar RM, Golbek TW, Kean KM, Brown WJ, Jana S, Baio JE, Karplus PA, and Mehl RA
- Subjects
- Carbonic Anhydrases chemistry, Carbonic Anhydrases metabolism, Cyclooctanes chemistry, Green Fluorescent Proteins metabolism, Humans, Models, Molecular, Immobilized Proteins metabolism
- Abstract
Biomaterials based on immobilized proteins are key elements of many biomedical and industrial technologies. However, applications are limited by an inability to precisely construct materials of high homogeneity and defined content. We present here a general "protein-limited immobilization" strategy by combining the rapid, bioorthogonal, and biocompatible properties of a tetrazine-strained trans -cyclooctene reaction with genetic code expansion to site-specifically place the tetrazine into a protein. For the first time, we use this strategy to immobilize defined amounts of oriented proteins onto beads and flat surfaces in under 5 min at submicromolar concentrations without compromising activity. This approach opens the door to generating and studying diverse protein-based biomaterials that are much more precisely defined and characterized, providing a greater ability to engineer properties across a wide range of applications.
- Published
- 2019
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