Helium beam shadowing for high spatial resolution patterning of antibodies on microstructured diagnostic surfaces

Eliedonna Cacao,Richard C Willson,Steven Kemper,Jennifer Knoop,Azeem Nasrullah,Paul Ruchhoeft,Robert L Atmar,Katerina Kourentzi,Tim Sherlock,Gila E Stein

doi:10.1186/1559-4106-8-9

Abstract

We have developed a technique for the high-resolution, self-aligning, and high-throughput patterning of antibody binding functionality on surfaces by selectively changing the reactivity of protein-coated surfaces in specific regions of a workpiece with a beam of energetic helium particles. The exposed areas are passivated with bovine serum albumin (BSA) and no longer bind the antigen. We demonstrate that patterns can be formed (1) by using a stencil mask with etched openings that forms a patterned exposure, or (2) by using angled exposure to cast shadows of existing raised microstructures on the surface to form self-aligned patterns. We demonstrate the efficacy of this process through the patterning of anti-lysozyme, anti-Norwalk virus, and anti-Escherichia coli antibodies and the subsequent detection of each of their targets by the enzyme-mediated formation of colored or silver deposits, and also by binding of gold nanoparticles. The process allows for the patterning of three-dimensional structures by inclining the sample relative to the beam so that the shadowed regions remain unaltered. We demonstrate that the resolution of the patterning process is of the order of hundreds of nanometers, and that the approach is well-suited for high throughput patterning.

Highlights

Creating patterned biological functionality of antibodies, enzymes, or cell-adhesion molecules is an essential tool for the development of high-performance bioanalytical devices and diagnostics
The image shows that the unexposed regions form silver deposits, as the antibodies remained functional, while the exposed regions were fully passivated by the bovine serum albumin (BSA)
In this study, we have demonstrated the use of a highthroughput collimated helium beam to pattern antibodycoated surfaces (1) through a stencil mask and (2) by using the geometry of existing three dimensional retroreflector structures to form self-aligned patterns

Summary

Introduction

Creating patterned biological functionality of antibodies, enzymes, or cell-adhesion molecules is an essential tool for the development of high-performance bioanalytical devices and diagnostics. Patterned antibody surfaces have previously been formed by ultraviolet (UV) [1,2,3] and electron beam [4,5,6] exposure of polymeric films, followed by a development step to create two chemicallydistinct surfaces which can be selectively functionalized. These approaches take advantage of well-established lithographic techniques and can achieve very high spatial resolution on planar substrates.

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Conclusion