Building three-dimensional nanostructures with active enzymes by surface templated layer-by-layer assembly

Abstract

We show that well-defined three-dimensional nanostructures of functional enzymes can be controllably fabricated by layer-by-layer assembly of avidin and biotinylated horseradish peroxidase on micro-contact printing patterned surface templates.

Nanoscale Biomolecular Structures on Self‐Assembled Monolayers Generated from Modular Pegylated Disulfides

“…[20] The work shown in this report will be useful in the future development of biomolecular "nanoarrays" for high throughput screening and wider applications in biomedical diagnostics, biosensors, organic material electronics and the assembly of more complex three-dimensional structures. [60] Nevertheless, the nanopatterned SAMs described can potentially find more immediate use in the investigation of cell biology mediated by cell surface interactions. These surfaces may also be a route to the isolation of single molecules in the study of single molecule dynamics and enzymology.…”

Preparation of bacteria microarray using selective patterning of polyelectrolyte multilayer and poly(ethylene glycol)-poly(lactide) diblock copolymer

“…21,22 Biomolecules including DNA, RNA, proteins, bacteria, and cell are easily immobilized in the PEL thin film by the combination of strong electrostatic interactions, hydrophobic interactions, and entrapment in the porous structure. [23][24][25][26] In this study, we have constructed a positively charged PEL surface at the outmost layer for the adhesion of bacteria because the net charge of bacteria in phosphate buffer (pH 7.5) is negative. 27,28 Most of cells have negatively charged surface because glycolipids and glycoproteins are located on the outer membrane.…”

“…The thickness effect on binding performance is well matched with the previous reports on a PEL coating for a protein microarray and cell microarray. [18][19][20][21][22][23][24][25][26][27][28][29] Bacterial Microarray. To explore patterns of bacteria on the functionalized surfaces, we have applied P. aeruginosa PAO1 that produces fluorescence signals due to expression of green fluorescence proteins based on cell population as a model bacterial organism.…”

Molecular Recognition and Specific Interactions for Biosensing Applications

“…We investigated surface engineering techniques using several SAMs (Figure 1A) such as 16-mercaptohexadecanoic acid (MHDA, a negatively charged carboxylic acid terminated thiol), 6-mercapto- N -hexylpyridinium bromide (MHP, a positively-charged pyridinium terminated thiol), 11-mercaptoundecyl hexa(ethylene glycol) alcohol (EG 6 OH, a neutral and flexible thiol terminated with hexa(ethylene glycol) group), and (2-[biotinamido]ethylamido)-3,3′-dithiodipropionic acid N -hydroxy-succinimide ester (biotin disulfide, a neutral and flexible disulfide group terminated with biotin) on gold substrate [3, 25]. Figure 1B shows the AFM topographic images of micropillars created by the LBL assembly of positively charged poly- l -lysine (PLL) and negatively charged poly(styrene-sulfonate) sodium salt (PSS) on microcontact printing (μCP)-patterned SAMs with 2 μm dot arrays of MHDA and backfilled with MHP on gold surface.…”

“…The functional surfaces provide ideal platforms for the immobilization of the desired biomolecules which can be achieved by physical adsorption, including electrostatic and hydrophobic interaction, covalent bonding, and specific interactions such as biotin-avidin, antibody-antigen interaction and DNA hybridization [2]. Especially the immobilization of biomolecules by the molecular recognition and specific interactions on the surfaces can yield good orientation and stability of the immobilized biomolecules thus leading to high-functionality within the biosensor fabrication process [3, 4]. …”