4.5 Article

CE methods for analysis of isoforms of prostate-specific antigen compatible with online derivatization for LIF detection

Journal

ELECTROPHORESIS
Volume 32, Issue 15, Pages 2036-2043

Publisher

WILEY-BLACKWELL
DOI: 10.1002/elps.201000524

Keywords

Biomarker; Glycoprotein; Glycosylation; Prostate cancer; PSA

Funding

  1. Spanish Ministry of Science and Innovation [CTQ2009-09399, PCI2006-A7-0646]
  2. Madrid Region Government [S-GEN-0247-2006]
  3. Spanish Ministry of Education

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Prostate-specific antigen (PSA) is the usual biomarker for prostate cancer (PCa). However, its lack of selectivity has lead to the search for new biomarkers. PSA glycosylation seems to depend on the pathophysiological conditions of the individual. Thus, methods to separate PSA isoforms (peaks) to study their role as PCa markers are needed. In this work, CE methods for PSA isoforms separation, based on the use of different dynamic coatings, are developed using UV detection. Three complementary CE methods allowing the separation of 8 or 9 PSA isoforms are selected. The longest method takes only 17 min, while the shortest one separates 9 isoforms in < 8 min. Depending on the isoforms of interest for their use as PCa biomarker, the CE method to be used can be chosen or various of them can be combined. A remarkable aspect of these methods is that the BGEs employed are devoid of compounds with primary amino groups, making the CE methods compatible with fluorescent on-column derivatization through amino residues. As a proof-of-concept, a preliminary result shows that LIF detection of labeled PSA analyzed by one of the three developed methods permits detection of glycoprotein isoforms.

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