Novel Protein Microarray Technology to Examine Men with Prostate Cancer
Abstract
The authors developed a novel macro and nanoporous silicon surface for protein microarrays to facilitate high-throughput biomarker discovery, and high-density protein-chip array analyses of complex biological samples. This surface offers 3D-surface enlarging properties and spot confinement, enabling both highly sensitive assays and high-density arrays with high precision. Relative standard deviation (RSD) of signal intensity both within and between chips was less than 20%. The detection limit and dynamic range of the novel protein chip surface was assessed with monoclonal anti-PSA IgG. Lower limit of detection was approximately equal to 0.7 ng/mL (26 pM) with linear detection (dynamic range) up to 104-fold higher levels. The authors also developed microfluid-nanovial arrays to enable rapid proteolytic digestion linked to matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. It uses minute amounts of sample and enhances both speed and accuracy in clinical applications of proteomics techniques, Automated picoliter microdispensing allows for precise fluid handling in the microarray format. To assess this platform in a putative clinical application, two prostate cancer biomarkers, PSA and human kallikrein 2 (hK2) were digested at levels of 100 fmol (PSA), 20 fmol (PSA), and 8 fmol (hK2). All biomarker digestions were completed in less than 30 seconds, with successful MS-identification in the porous nanovial setting.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 01, 2005
- Accession Number
- ADA457402
Entities
People
- Hans Lilja
Organizations
- Memorial Sloan Kettering Cancer Center