Development of proteomic tools to study protein adsorption on a biomaterial, titanium grafted with poly(sodium styrene sulfonate).
It is known that protein adsorption is the initial interaction between implanted biomaterials and biological environment. Generally, a complex protein layer will be formed on material surfaces within a few minutes and the composition of this layer at the interface determines the biological response to the implanted material, and therefore the long-term compatibility of the biomaterial. Despite different techniques exist to observe protein adsorption on biomaterials, none of them led to the identification of adsorbed proteins. In this paper, we report a chromatographic technique coupled to proteomics to analyse and identify proteins from complex biological samples adsorbed on biomaterial surfaces. This approach is based on (1) elaboration of the chromatographic support containing the biomaterial (2) a chromatography step involving adsorption of proteins on the biomaterial (3) the high-resolution separation of eluted proteins by 2-DE gel and (4) the identification of proteins by mass spectrometry. Experiments were performed with proteins from platelets rich plasma (PRP) adsorbed on a biomaterial which consist in titanium bioactivated with PolyNaSS. Our results show that chromatographic approach combined to 2-DE gels and mass spectrometry provides a powerful tool for the analysis and identification of proteins adsorbed on various surfaces.
Université Paris-UMR CNRS - CSPBAT - LBPS - UFR SMBH, Bobigny, France.
S
S
Oughlis
S
S
Lessim
S
S
Changotade
F
F
Bollotte
F
F
Poirier
G
G
Helary
J J
JJ
Lataillade
V
V
Migonney
D
D
Lutomski
D000327:Adsorption; D001672:Biocompatible Materials / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D013058:Mass Spectrometry; D008855:Microscopy, Electron, Scanning; D053657:Platelet-Rich Plasma / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013451:Sulfonic Acids / Q000737:chemistry*; D014025:Titanium / Q000737:chemistry*
D001672:Biocompatible Materials; D011108:Polymers; D011506:Proteins; D013451:Sulfonic Acids; D014025:Titanium; C077114:styrenesulfonic acid polymer