Indirect Microcontact Printing to Create Functional Patterns of Physisorbed Antibodies

Abstract

Microcontact printing (µCP) is a practical and versatile approach to create nanostructured patterns of biomolecular probes, but it involves conformational changes on the patterned bioreceptors that often lead to a loss on the biological activity of the resulting structures. Herein we introduce indirect µCP to create functional patterns of bioreceptors on solid substrates. This is a simple strategy that relies on physisorbing biomolecular probes of interest in the nanostructured gaps that result after patterning backfilling agents by standard µCP. This study presents the approach, assesses bovine serum albumin as backfilling agent for indirect µCP on different materials, reports the limitations of standard µCP on the functionality of patterned antibodies, and demonstrates the capabilities of indirect µCP to solve this issue. Bioreceptors were herein structured as diffractive gratings and used to measure biorecognition events in label-free conditions. Besides, as a preliminary approach towards sensing biomarkers, this work also reports the implementation of indirect µCP in an immunoassay to detect human immunoglobulin E.

BIO bragg gratings on microfibers for label-free biosensing

“…From these results, experimental detection and quantification limits of 0.1 µg mL −1 and 0.4 µg mL −1 , respectively, of IgG in label-free conditions are inferred. This is a promising sensitivity which is in the range of other recent label-free optical approaches (Chen et al, 2018;Gatterdam et al, 2017;Juste-Dolz et al, 2018;Makhneva et al, 2019).…”

“…This is an important and versatile technique in the state-of-art (Lamping et al, 2019;X. Wang et al, 2020), widely used to create functional and homogeneous patterns of biomolecules onto flat substrates of different compositions (Juste-Dolz et al, 2018). However, it remains challenging to pattern biomacromolecules onto curved, fragile, and micrometric structures as microfibers.…”

Lithographic Processes for the Scalable Fabrication of Micro- and Nanostructures for Biochips and Biosensors

“…Both direct and indirect strategies have been demonstrated in the fabrication of glass diffraction gratings for biosensing, where the grating consisted of stripes of antibody features 140 nm in width at a 555 nm pitch. 72 Here, the antibodies were printed either directly by μCP or indirectly by first printing bovine serum albumin by μCP and coating the unprinted areas with the antibody ( Figure 9 ). These methods therefore offered contrasting positive and negative tone patterning.…”

“…In indirect μCP, the first protein (red) is the backfilling, while the second protein (yellow) physisorbed is the probe, bound by the target. Redrawn by the authors from ref ( 72 ). (B,C,D) AFM topographic images of the different stages of indirect μCP.…”

Solid-Phase Microcontact Printing for Precise Patterning of Rough Surfaces: Using Polymer-Tethered Elastomeric Stamps for the Transfer of Reactive Silanes

“…Microstructured surfaces offer great potential in application fields such as microelectronics, 1 , 2 information storage, 3 , 4 (bio)sensing, 5 − 7 or optoelectronics. 8 , 9 In this context, microcontact printing (μCP) provides an inexpensive and straightforward access to surface patterning.…”