Abstract
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-1 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.
| Original language | English |
|---|---|
| Pages (from-to) | 1406-1409 |
| Number of pages | 4 |
| Journal | Biological and Pharmaceutical Bulletin |
| Volume | 23 |
| Issue number | 12 |
| Publication status | Published - 2000 |
| Externally published | Yes |
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Keywords
- Antioxidant
- Ceruloplasmin
- ESR spectroscopy
- Reactive oxygen specie
ASJC Scopus subject areas
- Molecular Medicine
- Pharmacology, Toxicology and Pharmaceutics(all)
Cite this
Purification of human ceruloplasmin as a by-product of C1-inhibitor. / Kouoh Elombo, Ferdinand; Radosevich, Miryana; Poulle, Michel; Descamps, Jacques; Chtourou, Sami; Burnouf, Thierry; Catteau, Jean Pierre; Bernier, Jean Luc; Cotelle, Nicole.
In: Biological and Pharmaceutical Bulletin, Vol. 23, No. 12, 2000, p. 1406-1409.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification of human ceruloplasmin as a by-product of C1-inhibitor
AU - Kouoh Elombo, Ferdinand
AU - Radosevich, Miryana
AU - Poulle, Michel
AU - Descamps, Jacques
AU - Chtourou, Sami
AU - Burnouf, Thierry
AU - Catteau, Jean Pierre
AU - Bernier, Jean Luc
AU - Cotelle, Nicole
PY - 2000
Y1 - 2000
N2 - 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-1 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.
AB - 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-1 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.
KW - Antioxidant
KW - Ceruloplasmin
KW - ESR spectroscopy
KW - Reactive oxygen specie
UR - http://www.scopus.com/inward/record.url?scp=0034537307&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034537307&partnerID=8YFLogxK
M3 - Article
C2 - 11145166
AN - SCOPUS:0034537307
VL - 23
SP - 1406
EP - 1409
JO - Biological and Pharmaceutical Bulletin
JF - Biological and Pharmaceutical Bulletin
SN - 0918-6158
IS - 12
ER -