lifesciences
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Miniaturization of multiplexed planar recombinant antibody arrays for serum protein profiling

Background

Antibody-based microarrays are a developing tool for high-throughput proteomics in health and disease. However, in order to enable global proteome profiling, novel miniaturized high-density antibody array formats must be developed.

Results

In this proof-of-concept study, we have designed a miniaturized planar recombinant (single-chain Fragment variable). antibody array technology platform for multiplexed profiling of non-fractionated, directly labelled serum samples. The size of the individual spot features was reduced 225-times (78.5 μm2/spot) and the array density was increased 19-times (38,000 spots/cm2). These miniaturized, multiplexed arrays were produced, using a desktop nanofabrication system based on dip-pen nanolithography technology, and interfaced with a high-resolution fluorescent-based scanner. The reproducibility, sensitivity, specificity, and applicability of the set-up were demonstrated by profiling a set of well-characterized serum samples.

Conclusion

The designed antibody array platform opens up new possibilities for large-scale, multiplex profiling of crude proteomes in a miniaturized fashion.

Credits:

Linn Petersson1, Michael Coen2, Nabil A Amro2, Lennart Truedsson3, Carl AK Borrebaek1 & Christer Wingren1

  • 1. Dept. Of Immunotechnology & CREATE Health, Lund University, Medicon Village, SE-22381 Lund, Sweden
  • 2. NanoInk, Inc., 8025 Lamon Ave. Skokie, IL 60077, USA
  • 3. Institute of Laboratory Medicine Section of Microbiology, Immunology & Glycobiology, Lund University, SE-22185 Lund, Sweden

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