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ACS Appl Mater Interfaces. 2019 Oct 9;11(40):36391-36398. doi: 10.1021/acsami.9b12746. Epub 2019 Sep 26.

Immobilization of Proteins with Controlled Load and Orientation.

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Department of Biochemistry and Biophysics , Oregon State University , 2011 Agricultural & Life Sciences Building , Corvallis , Oregon 97331-7305 , United States.
School of Chemical, Biological and Environmental Engineering , Oregon State University , 116 Johnson Hall, 105 SW 26th Street , Corvallis , Oregon 97331-7305 , United States.


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.


biointerface; biomaterial; bioorthogonal ligation; genetic code expansion; inverse electron demand Diels−Alder; protein immobilization; sTCO; tetrazine


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