Molecular Recognition-Mediated Fabrication of Protein Nanostructures by Dip-Pen Lithography

Abstract

We describe the molecular recognition-mediated, stepwise fabrication of patterned protein nanostructures with feature sizes on the order of 200 nm. First, a self-assembled monolayer (SAM) of 16-mercaptohexadecanoic acid (MHA) is patterned onto gold by dip-pen nanolithography (DPN), and the unpatterned regions are passivated with a protein-resistant oligoethylene glycol-terminated alkanethiol SAM. Next, an amine-terminated biotin derivative is covalently conjugated with the chemically activated MHA SAM nanopattern. The surface is then incubated with streptavidin to form streptavidin nanostructures, mediated by molecular recognition between biotin and streptavidin. Finally, protein nanopatterns are fabricated by molecular recognition-mediated immobilization of biotinylated protein from solution. Our fabrication methodology is generically applicable because of the ubiquity of biotin-tagged molecules.

Peptide-based biopolymers in biomedicine and biotechnology

“…In addition, bionanofabrication potentially allows direct interrogation of numerous biomolecules in a parallel manner. In materials science, molecular recognition of biomolecules such as DNA and antibodies can direct the assembly of nanoscale components such as gold nanoparticles, quantum dots, and proteins [23,[343][344][345][346]. Generation of more complex biomolecular ensembles is also possible when using self-assembling biomolecules.…”

Nanofabrication for Molecular Scale Devices

“…The water meniscus formed between the tip and the substrate by the condensation of humidity provides a transfer pathway by which the molecules are transferred from the tip to the substrate (Figure 7a). In (Hyun et al, 2002), the versatility of DPN was exploited in conjunction with other self-assembly methods. Specifically, 16-mercaptohexadecanoic acid was deposited onto gold by DPN and subsequently used as the grafting pattern for an amine-terminated biotin derivative.…”

Selective nanoshaving of self-assembled monolayers of 2-(4-pyridylethyl)triethoxysilane

“…Such examples are sometimes accompanied by replacement with another organic species [1,2,[8][9][10][11]. The former type of processing is nanoshaving [1][2][3][4][5][6][7] and the latter is "dip pen" nanolithography or "nanografting" [1,2,[8][9][10][11]. Selective oxidation of a self-assembled monolayer is also possible [2].…”

“…Selective oxidation of a self-assembled monolayer is also possible [2]. While this type of nanoshaving can lead to well defined "nanoscale" features in a monolayer molecular film [1][2][3][4][5][6][7][8][9][10][11], selective mechanical "cutting" or bond breaking within the molecule, for only part of the molecular film, has not been demonstrated. Over-cutting or etching using an AFM tip is certainly possible [3][4][5][6][7]12], but mechanically engineering the organic surface selectively within the monomolecular layer opens up new vistas for selective nanoscale surface modifications and surface chemistry.…”