Fabrication of selective anti-biofouling surface for micro/nanopatterning of proteins

Abstract

This paper reports a simple method for creating a functionalized surface for the efficient micro/nanopatterning of proteins by micromolding in capillaries (MIMIC) of poly(ethylene glycol)-poly(lactide) diblock copolymer (PEG-PLA) and self-assembled polyelectrolyte multilayers (PEL). The fabricated surface consisted of two distinct regions: a PEL region to promote protein immobilization and a PEG-PLA background as a biological barrier to prevent the nonspecific binding of proteins. When the ability of anti-biofouling of PEG-PLA was compared with the most widely used blocking agents, such as bovine serum albumin (BSA) and skim milk, the PEG-PLA prevents the nonspecific adsorption of several proteins. The properties of a functionalized surface were characterized by the water contact angle and atomic force microscopy (AFM). Topological analysis clearly indicated that the MIMIC method provides a reliable surface regardless of the micro-and nanopattern size. Two different functionalities of the fabricated surface produce uniform protein patterns from the micro-to nanoscale with a high signal to noise ratio. The proposed method allows for flexibility in forming shapes, such as lines, squares, circles, triangles and stars, and can control the pattern size from 400 nm to 90 µm. Finally, the antigen-antibody assay showed good linearity over the range of 10 ng/mL to 25 µg/mL, indicating its feasibility for a quantitative measurement of the concentration of target proteins in a sample.

Effect of Molecular Weight on the Capillary Absorption of Polymer Droplets

“…Indeed, there is a growing interest in building polymer-based functional nanomaterials and nanostructures for novel applications. − One of the methods used for producing these nanostructures is by filling nanomolds with the aid of capillary forces. For example, polymer-based nanowires, nanorods and nanotubes, nanoscale protein patterns, nanofibers, nanobelts, etc. are produced using capillary forces.…”

Multifunctional Structured Platforms: From Patterning of Polymer-Based Films to Their Subsequent Filling with Various Nanomaterials

“…The subsequent exposure of PEDOT:PSS films to water vapor induces swelling of the films followed by pattern replication driven by the capillary forces [ 140 ]. An interesting example where MCNIL was employed to fabricate arrays of submicrometer azopolymer pillars for unidirectional wetting and directional adhesion surfaces [ 103 ] ( Figure 6 h) was recently reported by Jo et al Other examples where MCNIL is employed include the fabrication of arrays of protein lines, squares, triangles, circles, or stars for selective anti-biofouling surfaces [ 141 ]. Yet other patterns obtained by MCNIL include protein dots perfectly aligned along grooves [ 132 ] or thermoplastic ( Figure 6 i) and conductive polymer wires [ 132 ].…”

Fabrication of protein micropatterns using a functional substrate with convertible protein‐adsorption surface properties

“…There are numerous reports of methods for successfully creating protein micropatterns on substrates using photolithography or microcontact printing techniques 3, 8–11. These methods are simple and enable the fabrication of micropatterns of a few proteins; however, it is difficult to create a complex protein micropattern composed of multiple proteins on a single substrate, as in a protein microarray.…”

“…Since the degree of spreading is influenced by the droplet conditions, including the additives and proteins contained in the droplets and the solvents of droplets, the printing size for each protein may differ, thus disturbing the creation of a precise and uniform protein pattern. Furthermore, the area outside the protein printed regions (background) in a protein microarray must be modified to resist nonspecific protein adsorption, which can prevent accurate examination of specific protein interactions 8. One approach that aims to overcome these drawbacks is the use of functional substrates that can be converted from a state that resists protein adsorption to a state that promotes protein adsorption in response to contact with specific solutions.…”