Immobilization of Proteins with Controlled Load and Orientation

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...

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Bibliographic Details
Published in:ACS applied materials & interfaces 2019-10, Vol.11 (40), p.36391-36398
Main Authors: Bednar, Riley M, Golbek, Thaddeus W, Kean, Kelsey M, Brown, Wesley J, Jana, Subhashis, Baio, Joe E, Karplus, P. Andrew, Mehl, Ryan A
Format: Article
Language:English
Subjects:
Carbonic Anhydrases - chemistry
Carbonic Anhydrases - metabolism
Cyclooctanes - chemistry
Green Fluorescent Proteins - metabolism
Humans
Immobilized Proteins - metabolism
Models, Molecular
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Summary: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.
ISSN:1944-8244
1944-8252
DOI:10.1021/acsami.9b12746