Proteomic analisys of protein extraction during hemofiltration with on-line endogenous reinfusion (HFR) using different polysulphone membranes
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.
KeywordsProtein Spot Continuous Renal Replacement Therapy Dialytic Treatment Uremic Toxin Ultra Filtrate
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.
- 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
- 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.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
- 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