Proteomic analisys of protein extraction during hemofiltration with on-line endogenous reinfusion (HFR) using different polysulphone membranes

  • Monari EmanuelaEmail author
  • Cuoghi Aurora
  • Bellei Elisa
  • Bergamini Stefania
  • Caiazzo Marialuisa
  • Aucella Filippo
  • Loschiavo Carmelo
  • Corazza Luca
  • Palladino Giuseppe
  • Sereni Luisa
  • Atti Mauro
  • Tomasi Aldo


In end-stage renal disease patients, extracorporeal dialytic therapy is not able to prevent the accumulation of toxins related to the uremic syndrome, a severe complication that increases morbidity and mortality rate. In this paper, hemoFiltration with on-line Reinfusion (HFR) architecture is used to evaluate the effect of a more permeable membrane on the extraction of medium–high molecular weight molecules. The aim of this study was to compare two polysulphone membranes for convective chamber: polyphenylene High Flux (pHF) and polyphenylene Super High-Flux (pSHF). Fourteen patients were subjected to HFR with pHF and pSHF membranes and ultra filtrate (UF) samples were collected to evaluate molecular weight cut-off (MWCO) and to identify extracted proteins. Furthermore, image analysis software was used in order to evaluate change in protein extraction during the dialysis. The quantification of four proteins by immunoassay demonstrates a higher permeability of pSHF membrane. Two-dimensional electrophoresis (2-DE) gels showed, for both membranes, the greater number of protein spots at 235 min. Some of the identified proteins, involved in nephropathic disease complications, were compared to assess differences in extraction during dialytic treatment by PDQuest analysis. UF proteomic analysis demonstrated a different behavior for the two membranes; pHF membrane was more permeable at the beginning of HFR treatment (15 min), while pSHF membrane at the end of treatment (235 min). Proteomic analysis is a suitable approach to investigate the behavior of different membranes during dialysis. Results indicated that pSHF membrane offers the higher permeability, and showed higher efficiency in removal of middle molecules related to uremic syndrome.


Protein Spot Continuous Renal Replacement Therapy Dialytic Treatment Uremic Toxin Ultra Filtrate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The “Fondazione Cassa di Risparmio di Modena” and Regione Emilia Romagna, (Italy) are acknowledged for financial support given for the acquisition of the MALDI-ToF/ToF. This study was partially financed by F.A.R.E. project (Filtration and Adsorption Regione Emilia Romagna). The authors thank Dr. Diego Pinetti and Dr. Daniela Manzini (C.I.G.S., University of Modena and Reggio Emilia, Italy) for the excellent technical support.

Conflict of interest

Caiazzo Marialuisa, Corazza Luca, Palladino Giuseppe, Sereni Luisa and Atti Mauro are Bellco srl. employees. The other authors declare no conflict of interest.

Supplementary material

10856_2014_5290_MOESM1_ESM.pdf (133 kb)
Supplementary material 1 (PDF 133 kb)
10856_2014_5290_MOESM2_ESM.pdf (1.8 mb)
Supplementary material 2 (PDF 1843 kb)


  1. 1.
    Dihazi H, Muller CA, Mattes H, Muller GA. Proteomic analysis to improve adequacy of hemo- and peritoneal dialysis: removal of small and high molecular weight proteins with high- and low-f lux filters or a peritoneal membrane. Proteomics Clin Appl. 2008;2:1167–82.CrossRefGoogle Scholar
  2. 2.
    Hussain S, Piering W, Mohyuddin T, Saleh M, Zhu Y-R, Hannan M, et al. Outcome among patients with acute renal failure needing continuous renal replacement therapy: a single center study. Hemodial Int. 2009;13:205–14.CrossRefGoogle Scholar
  3. 3.
    Vanholder R, Van Laecke S, Glorieux G. The middle-molecule hypothesis 30 years after: lost and rediscovered in the universe of uremic toxicity? J Nephrol. 2008;21:146–60.Google Scholar
  4. 4.
    Vanholder R, Baurmeister U, Brunet P, Cohen G, Glorieux G, Jankowski J. A bench to bedside view of uremic toxins. J Am Soc Nephrol. 2008;19:863–70.CrossRefGoogle Scholar
  5. 5.
    Vanholder R, De Smet R, Glorieux G, Argiles A, Baurmeister U, Brunet P, et al. Review on uremic toxins: classification, concentration, and interindividual variability. Kidney Int. 2003;63:1934–43.CrossRefGoogle Scholar
  6. 6.
    Duranton F, Cohen G, De Smet R, Rodriguez M, Jankowski J, Vanholder R, et al. Normal and pathologic concentrations of uremic toxins. J Am Soc Nephrol. 2012;23:1258–70.CrossRefGoogle Scholar
  7. 7.
    Galli F. Protein damage and inflammation in uraemia and dialysis patients. Nephrol Dial Transplant. 2007;22(Suppl 5):v20–36.CrossRefGoogle Scholar
  8. 8.
    Vanholder R, Meert N, Schepers E, Glorieux G. Uremic toxins: do we know enough to explain uremia? Blood Purif. 2008;26:77–81.CrossRefGoogle Scholar
  9. 9.
    Weissinger EM, Kaiser T, Meert N, De Smet R, Walden M, Mischak H, et al. Proteomics: a novel tool to unravel the patho-physiology of uraemia. Nephrol Dial Transplant. 2004;19:3068–77.CrossRefGoogle Scholar
  10. 10.
    Ghezzi PM, Botella J, Sartoris AM, Gervasio R, Diez C. Use of the ultrafiltrate obtained in two-chamber (PFD) hemodiafiltration as replacement fluid. Experimental ex vivo and in vitro study. Int J Artif Organs. 1991;14:327–34.Google Scholar
  11. 11.
    De Nitti C, Giordano R, Gervasio R, Castellano G, Podio V, Sereni L, et al. Choosing new adsorbents for endogenous ultrapure infusion fluid: performances, safety and flow distribution. Int J Artif Organs. 2001;24:765–76.Google Scholar
  12. 12.
    Tetta C, Ghezzi PM, De Nitti C, Fiorenzi A, Cianciavicchia D, Gervasio R. New options for on-line hemodiafiltration. Contrib Nephrol. 2002;137:212–20.CrossRefGoogle Scholar
  13. 13.
    Lou Wratten M, Ghezzi PM. Hemodiafiltration with endogenous reinfusion. Contrib Nephrol. 2007;158:94–102.CrossRefGoogle Scholar
  14. 14.
    Bolasco PG, Ghezzi PM, Serra A, Corazza L, Murtas S, Mascia M, et al. Hemodiafiltration with endogenous reinfusion with and without acetate-free dialysis solutions: effect on ESA requirement. Blood Purif. 2011;31:235–42.CrossRefGoogle Scholar
  15. 15.
    Bellei E, Bergamini S, Monari E, Fantoni LI, Cuoghi A, Ozben T, et al. High-abundance proteins depletion for serum proteomic analysis: concomitant removal of non-targeted proteins. Amino Acids. 2011;40:145–56.CrossRefGoogle Scholar
  16. 16.
    Bellei E, Rossi E, Lucchi L, Uggeri S, Albertazzi A, Tomasi A, et al. Proteomic analysis off early urinary biomarkers of renal changes in type 2 diabetic patients. Proteomics Clin Appl. 2008;2:478–91.CrossRefGoogle Scholar
  17. 17.
    Sirico ML, Guida B, Procino A, Pota A, Sodo M, Grandaliano G, et al. Human mature adipocytes express albumin and this expression is not regulated by inflammation. Mediat Inflamm. 2012;2012:236796.CrossRefGoogle Scholar
  18. 18.
    Armstrong JK, Wenby RB, Meiselman HJ, Fisher TC. The hydrodynamic radii of macromolecules and their effect on red blood cell aggregation. Biophys J. 2004;87:4259–70.CrossRefGoogle Scholar
  19. 19.
    Singer E, Markó L, Paragas N, Barasch J, Dragun D, Müller DN, et al. Neutrophil gelatinase-associated lipocalin: pathophysiology and clinical applications. Acta Physiol (Oxf). 2013;207:663–72.CrossRefGoogle Scholar
  20. 20.
    Musiał K, Zwolińska D. Neutrophil gelatinase-associated lipocalin (NGAL) and matrix metalloproteinases as novel stress markers in children and young adults on chronic dialysis. Cell Stress Chaperones. 2011;16:163–71.CrossRefGoogle Scholar
  21. 21.
    Le Quintrec M, Zuber J, Moulin B, Kamar N, Jablonski M, Lionet A, et al. Complement genes strongly predict recurrence and graft outcome in adult renal transplant recipients with atypical hemolytic and uremic syndrome. Am J Transplant. 2013;13:663–75.CrossRefGoogle Scholar
  22. 22.
    Wichman Z, Pajdak W, Smoleński O, Kopeć J. Detection of plasma proteins during sequential ultrafiltration/dialysis. Kidney Int. 1985;28:558–65.CrossRefGoogle Scholar
  23. 23.
    Jirka J, Burianova B, Jirka M, Blanicky P. Human serum Zn-alpha 2-glycoprotein in renal disease. Biomedicine. 1976;25:246–8.Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Monari Emanuela
    • 1
    Email author
  • Cuoghi Aurora
    • 1
  • Bellei Elisa
    • 1
  • Bergamini Stefania
    • 1
  • Caiazzo Marialuisa
    • 2
  • Aucella Filippo
    • 3
  • Loschiavo Carmelo
    • 4
  • Corazza Luca
    • 2
  • Palladino Giuseppe
    • 2
  • Sereni Luisa
    • 2
  • Atti Mauro
    • 2
  • Tomasi Aldo
    • 1
  1. 1.Department of Diagnostic, Clinical and Public Health MedicineUniversity of Modena and Reggio EmiliaModenaItaly
  2. 2.Scientific Affairs, Bellco s.r.lMirandolaItaly
  3. 3.IRCCS Hospital CSSSan Giovanni RotondoItaly
  4. 4.Division of Nephrology and DialysisLegnagoItaly

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