Author/Authors :
Niederkofler، Eric E. نويسنده , , Tubbs، Kemmons A. نويسنده , , Gruber، Karl نويسنده , , Nedelkov، Dobrin نويسنده , , Kiernan، Urban A. نويسنده , , Williams، Peter نويسنده , , Nelson، Randall W. نويسنده ,
Abstract :
A high-throughput mass spectrometric immunoassay (MSIA) system for the analysis of proteins directly from biological fluids is reported. A 96-well-format robotic workstation equipped with antibody-derivatized affinity pipet tips was used for the parallel extraction of specific proteins from samples and subsequent deposition onto 96-well arrayed matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) targets. Interferences from nonspecifically bound proteins were minimized through choice of appropriate affinity pipet tip derivatization chemistries. Sample preparation for MALDITOFMS was enhanced through the use of hydrophobic/hydrophilic contrasting targets, which also presented functionalities found to promote matrix/analyte crystal growth. Automated mass spectrometry was used in the unattended acquisition of data, resulting in an analysis rate of ~100 samples/h (biological fluid data). The quantitative MSIA of 2m levels present in human plasma samples is given as illustration.