MALDI-target integrated platform for affinity-captured protein digestion

• Main Authors: Ahmad Tajudin, Asilah, Adler, Belinda, Ekstrom, Simon, Marko-Varga, Gyorgy, Malm, Johan, Lilja, Hans, Laurell, Thomas
• Format: Article
• Published: Elsevier 2014
• Online Access: http://psasir.upm.edu.my/id/eprint/34429/; http://www.sciencedirect.com/science/article/pii/S0003267013011689

To address immunocapture of proteins in large cohorts of clinical samples high throughput sample processing is required. Here a method using the proteomic sample platform, ISET (integrated selective enrichment target) that integrates highly specific immunoaffinity capture of protein biomarker, digestion and sample cleanup with a direct interface to mass spectrometry is presented. The robustness of the on-ISET protein digestion protocol was validated by MALDI MS analysis of model proteins, ranging from 40 fmol to 1 pmol per nanovial. On-ISET digestion and MALDI MS/MS analysis of immunoaffinity captured disease-associated biomarker PSA (prostate specific antigen) from human seminal plasma are presented.