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Abstract: FR-PO482

The Proteome of Hemodialysis Membranes: A Discovery Proteomic Pilot Study

Session Information

Category: Dialysis

  • 701 Dialysis: Hemodialysis and Frequent Dialysis

Authors

  • Florens, Nans, Hospices Civils de Lyon, Lyon, France
  • Delolme, Frédéric, CNRS, Lyon, France
  • Juillard, Laurent, Hospices Civils de Lyon, Lyon, France
Background

Hemodialysis (HD) membranes have been continuously upgraded during the past decades. Nevertheless, little is known about the real pattern of protein removal among different HD membranes. We aimed to explore the proteome of depuration from a mid-cut-off HD membrane (Theranova, Baxter, IL, USA) and from a high flux membrane in HD and hemodiafiltration (HDF) (FX1000, Fresenius Medical Care, Bad Homburg, Germany).

Methods

9 HD patients were separated in 3 groups: Theranova, FX1000 in HD and FX1000 in HDF. During their mid-week session, 1 liter of dialysate was sampled and 30 ml were freeze-dried. Additionally, the dialysis membrane was eluted and both dialysate and eluate were prepared for LC/MS-MS analysis (liquid chromatography coupled with a tandem mass-spectrometer). Samples were analyzed using an nano-RSLC (high performance liquid chromatographer, Thermo Fischer, Waltham, MA, USA) coupled on line with a Q-Orbitrap mass spectrometer. Data were processed by database searching using SequestHT with Proteome Discoverer 2.2 software against a human Swissprot database and quantified with a TMT-labeling approach. Semi-quantitative analysis was expressed as a ratio. Proteins were analysed using STRING tool for reactome pathway analysis.

Results

526 proteins were found in the dialysate samples from all the membranes and 360 onto dialysis membranes. 455 proteins were found in the dialysate from FX1000 HDF group, 437 for Theranova and 410 for FX1000 HD. 360 proteins were also found adsorpbed onto the membranes. For Theranova, 45 proteins were found significantly more depurated by diffusion and/or convection than adsorption and 101 more by adsorption than diffusion/convection. For FX1000 HD, 56 and 100. For FX1000 HDF, 61 and 56 respectively. With Reactome pathway analysis, numerous removed proteins were involved in innate immune system, hemostasis, extra-cellular matrix organization, platelet aggregation, lipid metabolism and molecular signaling pathways.

Conclusion

More than 500 proteins were identified in the proteome of depuration from HD and HDF membranes among which a significant number are specific from each membrane and modalities. Further analysis are required to understand the issues of the depuration of these proteins in improving HD patients outcomes.

Funding

  • Commercial Support