Porous silicon surfaces: a candidate substrate for reverse protein arrays in cancer biomarker detection.
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 overproducing 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 mum and spot-to-spot center distances 100 mum, allowing microarray spot densities up to 14 000 spots per cm(2). 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.
Ressine
Anton
A
Department of Electrical Measurement, Lund University, Lund, Sweden.
Corin
Irina
I
Järås
Kerstin
K
Guanti
Ginevra
G
Simone
Cristiano
C
Marko-Varga
György
G
Laurell
Thomas
T
D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western / Q000379:methods; D002457:Cell Extracts / Q000032:analysis; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000737:chemistry* / Q000175:diagnosis / Q000378:metabolism; D016213:Cyclins / Q000032:analysis; D019085:Fluorescent Antibody Technique, Direct; D045325:HCT116 Cells; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry*; D013499:Surface Properties; D046888:Tissue Array Analysis / Q000379:methods*; D014020:Tissue Extracts / Q000032:analysis / Q000737:chemistry
D014408:Biomarkers, Tumor; D002457:Cell Extracts; D016213:Cyclins; D009363:Neoplasm Proteins; D014020:Tissue Extracts; D012825:Silicon