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Submitted on February 12, 2005
Revised on February 20, 2005
Accepted on February 20, 2005

A proteomic analysis of human hemodialysis fluid

Henrik Molina, Jakob Bunkenborg, G. Hanumanthu Reddy, Babylakshmi Muthusami, Paul J Scheel, and Akhilesh Pandey

McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, MD 21205

Corresponding Author: pandey{at}jhmi.edu

The vascular compartment is an easily accessible compartment that provides an opportunity to measure analytes for diagnostic, prognostic or therapeutic indications. Both serum and plasma have been analyzed extensively by proteomic approaches in an effort to catalog all proteins and polypeptides. Limitations of such approaches in obtaining a comprehensive catalog of proteins include the fact that a handful of proteins constitute over 90 percent of plasma protein content and that the renal glomeruli filter out proteins and polypeptides that are smaller than 66 kDa from blood. We chose to study hemodialysis fluid because it contains a higher concentration of small proteins and polypeptides and is also simultaneously depleted for the most abundant proteins present in blood. Using gel electrophoresis in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS), we identified 292 proteins, of which greater than 70 percent had not been previously identified from serum or plasma. More than half of the proteins identified from the hemodialysis fluid were smaller than 40 kDa. We also found 50 N-terminally acetylated peptides that allowed us to unambiguously map the N-termini of mature forms of the corresponding proteins. Several identified proteins, including cytokines, were only present as predicted transcripts in databases and thus represent novel proteins. The proteins identified in this study could serve as biomarkers in serum using more sensitive methods such as ELISA specific antibodies.


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