Purification of human ceruloplasmin as a by-product of C1-inhibitor.
Human ceruloplasmin (Cp) has been purified from cryoprecipitate-poor plasma as a by-product of the C1-inhibitor production chain. Highly purified Cp was obtained by subsequent ion-exchange chromatography on sulfate-Fractogel EMD and TMAE-Fractogel EMD. Treatments for viral safety included application of the solvent-detergent method and two nanofiltration steps using 35- and 15-nm pore size filters at the end of the process. Overall antigen yield was 95 (+/-5) %. Purified human ceruloplasmin was studied by electron spin resonance (ESR) to characterize its different types of copper complexes and to check its antioxidant properties. We distinguished three types of complexes: one type-2 Cu(II) with g// = 2.25 and A// = 180 G and two type-I Cu(II) exhibiting different narrow hyperfine splitting (A// = 72 G and A// = 90 G) with close g// (2.20 and 2.21). Purified Cp has a specific activity of 24.5+/-0.2 mU/mg of proteins. This process provides a method for Cp purification that could be easily integrated into modern plasma fractionation.
Kouoh Elombo
F
F
Faculté des Sciences Pharmaceutiques et Biologiques, Laboratoire de Pharmacologie, Pharmacocinétique et Pharmacie Clinique, Lille, France. elferdy@caramail.com
Radosevich
M
M
Poulle
M
M
Descamps
J
J
Chtourou
S
S
Burnouf
T
T
Catteau
J P
JP
Bernier
J L
JL
Cotelle
N
N
D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002570:Ceruloplasmin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods; D003174:Complement C1 Inactivator Proteins / Q000096:biosynthesis*; D050718:Complement C1 Inhibitor Protein; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D005374:Filtration; D006801:Humans; D007122:Immunoelectrophoresis; D017382:Reactive Oxygen Species / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D019820:Xanthine / Q000737:chemistry / Q000378:metabolism; D014969:Xanthine Oxidase / Q000737:chemistry / Q000378:metabolism
D000975:Antioxidants; D003174:Complement C1 Inactivator Proteins; D050718:Complement C1 Inhibitor Protein; D017382:Reactive Oxygen Species; D019820:Xanthine; D002570:Ceruloplasmin; D014969:Xanthine Oxidase