Patterned Protein Microarrays for Bacterial Detection

Abstract

Patterned microarrays of antibodies were fabricated and tested for their ability to bind targeted bacteria. These arrays were used in a series of bacterial immunoassays to detect E. coli 0157:H7 and Renibacterium salmoninarum (RS). Microarrays were fabricated using microcontact printing (µCP) and characterized using scanning probe microscopy (SPM). The high-resolution SPM imaging showed that targeted bacteria had a higher binding selectivity to complementary antibody patterns than to unfunctionalized regions of the substrate. Additional studies indicated a significant reduction in binding of bacteria when the microarrays were exposed to nonspecific bacteria. These studies demonstrate how protein microarrays could be developed into useful platforms for sensing microorganisms.

Irreversible adsorption of particles on heterogeneous surfaces

“…By using the alternative layer-by-layer deposition method involving polyelectrolytes, multilayer particle films of a targeted architecture can be produced [9 -13]. In biomedical applications special proteins (antibodies) attached to the surface are used for a selective binding of a desired ligands from protein mixtures as is the case of the affinity chromatography [14], recognition processes (biosensors) [15,16], immunological assays [17,18], etc.…”

Electron beam and mechanical lithographies as enabling factors for organic-based device fabrication

“…1), which was originally proposed for the patterning of self-assembled monolayers alkanethiolates onto gold [12], and soon applied to the delivery of patterned proteins onto surfaces [10,13]. In the last years, the capability of producing patterns of biological molecules with micrometer-level resolution has attracted an increasing interest, in view of their possible use in a wide number of applications, such as microarray technologies for genomics and proteomics, medical diagnostics, biological assays and sensors [14], molecular electronics [15,16], control of the cellular adhesion, growth and functionality [17], and bacterial detection [18]. The application of the microcontact printing to both extracellular matrix networks and metalloproteins shows an excellent pattern definition and chemical contrast (Fig.…”

Probing interfacial interactions of bacteria on metal nanoparticles and substrates with different surface properties

“…Despite increasing reports on AFM imaging of bacteria on surfaces, little is known about the interfacial interactions. The use of microcontact printing techniques [23,24] to pattern proteins onto substrates without loss of biological activity [25][26][27] is one of the effective ways reported for developing an understanding of interfacial interactions. For example, the use of polymer/alkanethiol-patterned substrates was demonstrated for AFM imaging of bacteria, in which the n-alkanethiol wells promote bacterial adhesion whereas walls consisting of hydrophilic poly(acrylic acid)/poly(ethylene glycol) layered nanocomposites inhibit bacterial adhesion [28].…”