Porous silicon surfaces - A candidate substrate for reverse protein arrays in cancer biomarker detection

Anton Ressine, Irina Corin, Kerstin Järås, Ginevra Guanti, Cristiano Simone, György Marko-Varga, Thomas Laurell

Research output: Contribution to journalArticlepeer-review

Abstract

This paper introduces a new substrate for reverse-phase protein microarray applications based on macroporous silicon. A key feature of the microarray substrate is the vastly surface enlarging properties of the porous silicon, which simultaneously offers highly confined microarray spots. The proof of principle of the reverse array concept was demonstrated in the detection of different levels of cyclin E, a possible cancer biomarker candidate which regulates G1-S transition and correlates with poor prognosis in different types of human cancers. The substrate properties were studied performing analysis of total cyclin E expression in human colon cancer cell lines Hct116 and SW480. The absence of unspecific binding and good microarray quality was demonstrated. In order to verify the performance of the 3-D textured macroporous surface for complex biological samples, lysates of the human tissue spiked to different levels with cell extract over-producing cyclin E (Hct116) were arrayed on the chip surface. The samples were spotted in a noncontact mode in 100 pL droplets with spots sizes ranged between 50 and 70 μm and spot-to-spot center distances 100 μm, allowing microarray spot densities up to 14 000 spots per cm2. The different sample types of increasing complexities did not have any impact on the spot intensities recorded and the protein spots showed good homogeneity and reproducibility over the recorded microarrays. The data demonstrate the potential use of macroporous silicon as a substrate for quantitative determination of a cancer biomarker cyclin E in tissue lysates.

Original languageEnglish
Pages (from-to)4407-4415
Number of pages9
JournalElectrophoresis
Volume28
Issue number23
DOIs
Publication statusPublished - Dec 2007

Keywords

  • Biomarker detection
  • Porous silicon
  • Reverse lysate arrays

ASJC Scopus subject areas

  • Clinical Biochemistry

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