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Co‐Immobilization of Proteins and DNA Origami Nanoplates to Produce High‐Contrast Biomolecular Nanoarrays
Author(s) -
Hager Roland,
Burns Jonathan R.,
Grydlik Martyna J.,
Halilovic Alma,
Haselgrübler Thomas,
Schäffler Friedrich,
Howorka Stefan
Publication year - 2016
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201600311
Subject(s) - dna origami , nanobiotechnology , nanotechnology , biosensor , dna , materials science , substrate (aquarium) , fluorescence , dna nanotechnology , nanostructure , biomolecule , chemistry , biophysics , nanoparticle , biology , biochemistry , ecology , physics , quantum mechanics
The biofunctionalization of nanopatterned surfaces with DNA origami nanostructures is an important topic in nanobiotechnology. An unexplored challenge is, however, to co‐immobilize proteins with DNA origami at pre‐determined substrate sites in high contrast relative to the nontarget areas. The immobilization should, in addition, preferably be achieved on a transparent substrate to allow ultrasensitive optical detection. If successful, specific co‐binding would be a step towards stoichiometrically defined arrays with few to individual protein molecules per site. Here, we successfully immobilize with high specificity positively charged avidin proteins and negatively charged DNA origami nanoplates on 100 nm‐wide carbon nanoislands while suppressing undesired adsorption to surrounding nontarget areas. The arrays on glass slides achieve unprecedented selectivity factors of up to 4000 and allow ultrasensitive fluorescence read‐out. The co‐immobilization onto the nanoislands leads to layered biomolecular architectures, which are functional because bound DNA origami influences the number of capturing sites on the nanopatches for other proteins. The novel hybrid DNA origami‐protein nanoarrays allow the fabrication of versatile research platforms for applications in biosensing, biophysics, and cell biology, and, in addition, represent an important step towards single‐molecule protein arrays.