Protein-adsorptive properties are a key feature of membranes used for haemodialysis treatment.Protein adsorption is vital to the biocompatibility of a membrane material and influences membrane’sperformance. The object of the present study is to investigate membrane biocompatibility bycorrelating the adsorbed proteome repertoire with chemical feature of the membrane surfaces.Dialyzers composed of either cellulose triacetate (Sureflux 50 L, effective surface area 0.5 m2; NiproCorporation, Japan) or the polysulfone-based helixone (FX40, effective surface area 0.4 m2; FreseniusMedical Care AG, Germany) materials were employed to develop an ex vivo apparatus to studyprotein adsorption. Adsorbed proteins were eluted by a strong chaotropic buffer condition andinvestigated by a proteomic approach. The profiling strategy was based on 2D-electrophoresisseparation of desorbed protein coupled to MALDI-TOF/TOF analysis. The total protein adsorptionwas not significantly different between the two materials. An average of 179 protein spots wasvisualised for helixone membranes while a map of retained proteins of cellulose triacetate membraneswas made up of 239 protein spots. The cellulose triacetate material showed a higher binding capacityfor albumin and apolipoprotein. In fact, a number of different protein spots belonging to the genetranscript of albumin were visible in the cellulose triacetate map. In contrast, helixone bound only asmall proportion of albumin, while proved to be particularly active in retaining protein associatedwith the coagulation cascade, such as the fibrinogen isoforms. Our data indicate that proteomictechniques are a useful approach for the investigation of proteins surface-adsorbed ontohaemodialysis membranes, and may provide a molecular base for the interpretation of the efficacyand safety of anticoagulation treatment during renal replacement therapy.
Protein-adsorptive properties are a key feature of membranes used for haemodialysis treatment. Protein adsorption is vital to the biocompatibility of a membrane material and influences membrane's performance. The object of the present study is to investigate membrane biocompatibility by correlating the adsorbed proteome repertoire with chemical feature of the membrane surfaces. Dialyzers composed of either cellulose triacetate (Sureflux 50 L, effective surface area 0.5 m(2); Nipro Corporation, Japan) or the polysulfone-based helixone (FX40, effective surface area 0.4 m(2); Fresenius Medical Care AG, Germany) materials were employed to develop an ex vivo apparatus to study protein adsorption. Adsorbed proteins were eluted by a strong chaotropic buffer condition and investigated by a proteomic approach. The profiling strategy was based on 2D-electrophoresis separation of desorbed protein coupled to MALDI-TOF/TOF analysis. The total protein adsorption was not significantly different between the two materials. An average of 179 protein spots was visualised for helixone membranes while a map of retained proteins of cellulose triacetate membranes was made up of 239 protein spots. The cellulose triacetate material showed a higher binding capacity for albumin and apolipoprotein. In fact, a number of different protein spots belonging to the gene transcript of albumin were visible in the cellulose triacetate map. In contrast, helixone bound only a small proportion of albumin, while proved to be particularly active in retaining protein associated with the coagulation cascade, such as the fibrinogen isoforms. Our data indicate that proteomic techniques are a useful approach for the investigation of proteins surface-adsorbed onto haemodialysis membranes, and may provide a molecular base for the interpretation of the efficacy and safety of anticoagulation treatment during renal replacement therapy.
Proteomic analysis of protein adsorption capacity of different haemodialysis membranes
Pieroni Luisa;
2012-01-01
Abstract
Protein-adsorptive properties are a key feature of membranes used for haemodialysis treatment. Protein adsorption is vital to the biocompatibility of a membrane material and influences membrane's performance. The object of the present study is to investigate membrane biocompatibility by correlating the adsorbed proteome repertoire with chemical feature of the membrane surfaces. Dialyzers composed of either cellulose triacetate (Sureflux 50 L, effective surface area 0.5 m(2); Nipro Corporation, Japan) or the polysulfone-based helixone (FX40, effective surface area 0.4 m(2); Fresenius Medical Care AG, Germany) materials were employed to develop an ex vivo apparatus to study protein adsorption. Adsorbed proteins were eluted by a strong chaotropic buffer condition and investigated by a proteomic approach. The profiling strategy was based on 2D-electrophoresis separation of desorbed protein coupled to MALDI-TOF/TOF analysis. The total protein adsorption was not significantly different between the two materials. An average of 179 protein spots was visualised for helixone membranes while a map of retained proteins of cellulose triacetate membranes was made up of 239 protein spots. The cellulose triacetate material showed a higher binding capacity for albumin and apolipoprotein. In fact, a number of different protein spots belonging to the gene transcript of albumin were visible in the cellulose triacetate map. In contrast, helixone bound only a small proportion of albumin, while proved to be particularly active in retaining protein associated with the coagulation cascade, such as the fibrinogen isoforms. Our data indicate that proteomic techniques are a useful approach for the investigation of proteins surface-adsorbed onto haemodialysis membranes, and may provide a molecular base for the interpretation of the efficacy and safety of anticoagulation treatment during renal replacement therapy.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
