Abstract
Depositing biomolecule micropatterns on solid substrates via microcontact printing (µCP) usually requires complex chemical substrate modifications to initially create reactive surface groups. Here, we present a simplified activation procedure for untreated solid substrates based on a commercial polymer metal ion coating (AnteoBindTM Biosensor reagent) that allows for direct µCP and the strong attachment of proteins via avidity binding. In proof-of-concept experiments, we identified the optimum working concentrations of the surface coating, characterized the specificity of protein binding and demonstrated the suitability of this approach by subcellular micropatterning experiments in living cells. Altogether, this method represents a significant enhancement and simplification of existing µCP procedures and further increases the accessibility of protein micropatterning for cell biological research questions.
Highlights
Depositing biomolecules onto solid substrates in regular 2D patterns with micrometer resolution, known as molecular printing or, more commonly, as protein micropatterning, has found widespread use in academic laboratories, and multiple applications for biomedical and cell biological research have emerged [1,2,3,4,5,6,7,8]
Many different methodologies have been developed in the recent years, whereas the fabrication of biomolecule micropatterned solid substrates is mainly depending on the intended application and available lab infrastructure
Direct protein deposition can be realized by approaches such as dip-pen lithography (DPL) [14], microchannel cantilever spotting [15], polymer pen lithography (PPL) [16], microfluidic patterning [17], fluidic force microscopy (FluidFM) [18] and soft lithography via microcontact printing [19]
Summary
Depositing biomolecules onto solid substrates in regular 2D patterns with micrometer resolution, known as molecular printing or, more commonly, as protein micropatterning, has found widespread use in academic laboratories, and multiple applications for biomedical and cell biological research have emerged [1,2,3,4,5,6,7,8] In this regard, many different methodologies have been developed in the recent years, whereas the fabrication of biomolecule micropatterned (and even nanopatterned) solid substrates is mainly depending on the intended application and available lab infrastructure. Protein patterning by μCP provides unique features compared with all other sophisticated patterning technologies as it is: (i) highly reproducible and robust, (ii) easy to perform, (iii) extremely fast, (iv) modularly expandable (with respect to feature size and printed protein), (v) comparatively cheap, and (vi) independent of special lab equipment
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