Fabrication and imaging of two-dimensional patterns of proteins adsorbed on self-assembled monolayers by scanning electron microscopy

López, Gabriel P.; Biebuyck, Hans A.; Härter, Ralph; Kumar, Amit; Whitesides, George M.

doi:10.1021/ja00076a038

Cited by 194 publications

(182 citation statements)

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“…21,22 These were then patterned with nickel (II)-chelating nitrilotriacetic acid (Ni-NTA) terminated alkyl thiols (Fig. 1a) using two-step nanografting 13,15 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Physical Controls on Directed Virus Assembly at Nanoscale Chemical Templates

et al. 2006

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Viruses are attractive building blocks for nanoscale heterostructures, but little is understood about the physical principles governing their directed assembly. In-situ force microscopy was used to investigate organization of Cowpea Mosaic Virus engineered to bind specifically and reversibly at nanoscale chemical templates with sub-30nm features. Morphological evolution and assembly kinetics were measured as virus flux and inter-viral potential were varied. The resulting morphologies were similar to those of atomic-scale epitaxial systems, but the underlying thermodynamics was analogous to that of colloidal systems in confined geometries. The 1D templates biased the location of initial cluster formation, introduced asymmetric sticking probabilities, and drove 1D and 2D condensation at subcritical volume fractions. The growth kinetics followed a t 1/2 law controlled by the slow diffusion of viruses. The lateral expansion of virus clusters that initially form on the 1D templates following introduction of polyethylene glycol (PEG) into the solution suggests a significant role for weak

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“…21,22 These were then patterned with nickel (II)-chelating nitrilotriacetic acid (Ni-NTA) terminated alkyl thiols (Fig. 1a) using two-step nanografting 13,15 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Physical Controls on Directed Virus Assembly at Nanoscale Chemical Templates

et al. 2006

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“…Without using templates, DNA micropatterns may be directly produced by using photolithography (12)(13)(14). Micropatterns of proteins have also been created by first making patterned surfaces as templates, followed by selective adsorption of proteins (15). More precise positioning of biomolecules requires new fabrication strategies.…”

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confidence: 99%

Positioning protein molecules on surfaces: A nanoengineering approach to supramolecular chemistry

Liu

Amro

2002

Proc. Natl. Acad. Sci. U.S.A.

183

158

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We discuss a nanoengineering approach for supramolecular chemistry and self assembly. The collective properties and biofunctionalities of molecular ensembles depend not only on individual molecular building blocks but also on organization at the molecular or nanoscopic level. Complementary to ''bottom-up'' approaches, which construct supramolecular ensembles by the design and synthesis of functionalized small molecular units or large molecular motifs, nanofabrication explores whether individual units, such as small molecular ligands, or large molecules, such as proteins, can be positioned with nanometer precision. The separation and local environment can be engineered to control subsequent intermolecular interactions. Feature sizes as small as 2 ؋ 4 nm 2 (32 alkanethiol molecules) are produced. Proteins may be aligned along a 10-nm-wide line or within two-dimensional islands of desired geometry. These high-resolution engineering and imaging studies provide new and molecular-level insight into supramolecular chemistry and self-assembly processes in bioscience that are otherwise unobtainable, e.g., the influence of size, separation, orientation, and local environment of reaction sites. This nanofabrication methodology also offers a new strategy in construction of two-and three-dimensional supramolecular structures for cell, virus, and bacterial adhesion, as well as biomaterial and biodevice engineering. S upramolecular chemistry has become an area of intense research (1-4), partly inspired by biological ensembles in nature, such as collagen and enzymes or protein assemblies in general. In nature, the collective properties and biofunctionalities of these ensembles depend not only on the individual molecular units but also (perhaps even more importantly) on the organization at the molecular or nanoscopic level (1, 2, 4, 5). Such organization dependence can be attributed to polyvalent interactions in biological systems (6). ''Bottom-up'' approaches have been taken to mimic nature and have resulted in creative synthesis of small molecular units (7,8) and large molecular motifs (9). These molecular building blocks contain the desired charge, polarization, or chemical functionalities that will affect intermolecular interactions such as van der Waals forces, hydrogen bonding, polar attractions, and͞or hydrophobic interactions (7-9). These interactions dictate the subsequent assembly into supramolecular structures (4, 9-11). Complementary to these synthetic approaches, we explore whether individual units such as small molecular ligands or large molecules such as proteins can be positioned with nanometer precision by using nanoengineering methodologies. The separation and local environment can be engineered to influence subsequent intermolecular interactions.Micrometer-sized patterns of biomolecules can be produced by using relatively well known microfabrication technologies. Without using templates, DNA micropatterns may be directly produced by using photolithography (12)(13)(14). Micropatterns of proteins have also been ...

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“…SAMs are increasingly important tools in the micro-and nanolithographic patterning of organic and inorganic materials. They have recently been used for selected area polypeptide synthesis (1), as barriers to chemical etching (2,3), for the formation of microcrystals (4,5) or microelectrodes (4), for creation of electrically conducting polypyrrole circuitry (6), for patterned water condensation (4), and for adhesion of cells (7)(8)(9)(10) and proteins (8,(11)(12)(13)(14)(15)(16)(17). OTMS is one of the most widely used of the alkyl silanes for producing hydrophobic SAMs on SiO2.…”

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confidence: 99%

Patterning of functional antibodies and other proteins by photolithography of silane monolayers.

Mooney

Hunt

McIntosh

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

248

165

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We have demonstrated the assembly of twodimensional patterns of functional antibodies on a surface. In particular, we have selectively adsorbed micrometer-scale regions of biotinylated immunoglobulin that exhibit specific antigen binding after adsorption. The advantage of this technique is its potential adaptability to adsorbing arbitrary proteins in tightly packed monolayers while retaining functionality. The procedure begins with the formation of a self-assembled monolayer of n-octadecyltrimethoxysilane (OTMS) on a silicon dioxide surface. This monolayer can then be selectively removed by UV photolithography. Under appropriate solution conditions, the OTMS regions will adsorb a monolayer of bovine serum albumin (BSA), while the silicon dioxide regions where the OTMS has been removed by UV light will adsorb less than 2% of a monolayer, thus creating high contrast patterned adsorption of BSA. The attachment of the molecule biotin to the BSA allows the pattern to be replicated in a layer of streptavidin, which bonds to the biotinylated BSA and in turn will bond an additional layer of an arbitrary biotinylated protein. In our test case, functionality of the biotinylated goat antibodies raised against mouse immunoglobulin was demonstrated by the specific binding of fluorescently labeled mouse IgG.We are interested in the development and application of techniques for protein patterning, especially for in vitro studies of protein function and for applications outside of the biological context (e.g., for artificial biomineralization and in biochemical sensors). In this paper we describe a general technique for producing two-dimensional patterns of functional protein on silicon dioxide surfaces by using a combination of self-assembled monolayers (SAMs) of alkyl silanes, albuminalkyl adsorption, and biotin-avidin interactions. We illustrate the technique by generating two-dimensional patterns of functional goat antibodies raised against mouse immunoglobulins (GAM).Our procedure begins with the formation of an noctadecyltrimethoxysilane (OTMS) ¶ SAM on a sio2 substrate. SAMs are increasingly important tools in the micro-and nanolithographic patterning of organic and inorganic materials. They have recently been used for selected area polypeptide synthesis (1), as barriers to chemical etching (2, 3), for the formation of microcrystals (4, 5) or microelectrodes (4), for creation of electrically conducting polypyrrole circuitry (6), for patterned water condensation (4), and for adhesion of cells (7-10) and proteins (8,(11)(12)(13)(14)(15)(16)(17). OTMS is one of the most widely used of the alkyl silanes for producing hydrophobic SAMs on SiO2. The SAM can be selectively removed from the surface by ultraviolet (UV) exposure through a lithographic mask. The difference in adsorption of bovine serum albumin (BSA) to the exposed and unexposed regions of the surface creates a two-dimensional pattern of BSA with a surface coverage "50 times greater on the OTMS than on the exposed sio2. If the BSA is tagged with biotin, the p...

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Fabrication and imaging of two-dimensional patterns of proteins adsorbed on self-assembled monolayers by scanning electron microscopy

Cited by 194 publications

References 0 publications

Physical Controls on Directed Virus Assembly at Nanoscale Chemical Templates

Physical Controls on Directed Virus Assembly at Nanoscale Chemical Templates

Positioning protein molecules on surfaces: A nanoengineering approach to supramolecular chemistry

Patterning of functional antibodies and other proteins by photolithography of silane monolayers.

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