Large-scale purification and characterization of non-glycosylated Gc globulin (vitamin D-binding protein) from plasma fraction IV. 2006

Charlotte S Jørgensen, and Maja Christiansen, and Inga Laursen, and Lisbeth B Krogsøe, and Peter Højrup, and Lene Blou, and Gunnar Houen
Department of Research and Development, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen, Denmark. csv@ssi.dk

Gc globulin, also called vitamin D-binding protein, is a plasma protein involved in the extracellular actin-scavenger system. Low levels of Gc globulin have been found to correlate with multiple organ failure and nonsurvival of patients with fulminant hepatic failure and trauma. Therefore substitution therapy with Gc globulin might be beneficial for such patients, increasing their chance of survival. In the present study, we describe a large-scale purification process for the production of a virus-safe human plasma-derived Gc globulin from Cohn fraction IV paste. The process includes three ion-exchange-chromatography steps, followed by a gel filtration, and two virus-reduction steps are implemented. The Gc globulin product was characterized with respect to purity, functional activity, glycosylation and, finally, with respect to content of endotoxin. From the results, it can be concluded that human Gc globulin purified from Cohn fraction IV is non-glycosylated. The purified Gc globulin is able to mask the presence of endotoxin by 20%.

UI MeSH Term Description Entries
D009102 Multiple Organ Failure A progressive condition usually characterized by combined failure of several organs such as the lungs, liver, kidney, along with some clotting mechanisms, usually postinjury or postoperative. MODS,Multiple Organ Dysfunction Syndrome,Organ Dysfunction Syndrome, Multiple,Organ Failure, Multiple,Failure, Multiple Organ,Multiple Organ Failures
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D002852 Chromatography, Ion Exchange Separation technique in which the stationary phase consists of ion exchange resins. The resins contain loosely held small ions that easily exchange places with other small ions of like charge present in solutions washed over the resins. Chromatography, Ion-Exchange,Ion-Exchange Chromatography,Chromatographies, Ion Exchange,Chromatographies, Ion-Exchange,Ion Exchange Chromatographies,Ion Exchange Chromatography,Ion-Exchange Chromatographies
D004586 Electrophoresis An electrochemical process in which macromolecules or colloidal particles with a net electric charge migrate in a solution under the influence of an electric current. Electrophoreses
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D014809 Vitamin D-Binding Protein An alpha-globulin found in the plasma of man and other vertebrates. It is apparently synthesized in the liver and carries vitamin D and its metabolites through the circulation and mediates the response of tissue. It is also known as group-specific component (Gc). Gc subtypes are used to determine specific phenotypes and gene frequencies. These data are employed in the classification of population groups, paternity investigations, and in forensic medicine. Group-Specific Component Globulin,Vitamin D-Binding Globulin,25-HCC-Binding Protein,25-Hydroxycholecalciferol-Binding Protein,25-Hydroxyvitamin D-Binding Protein,Calciferol-Binding Protein,Gc Globulin,Transcalciferin,25 HCC Binding Protein,25 Hydroxycholecalciferol Binding Protein,25 Hydroxyvitamin D Binding Protein,Calciferol Binding Protein,Component Globulin, Group-Specific,Globulin, Gc,Globulin, Group-Specific Component,Globulin, Vitamin D-Binding,Group Specific Component Globulin,Vitamin D Binding Globulin,Vitamin D Binding Protein
D017114 Liver Failure, Acute A form of rapid-onset LIVER FAILURE, also known as fulminant hepatic failure, caused by severe liver injury or massive loss of HEPATOCYTES. It is characterized by sudden development of liver dysfunction and JAUNDICE. Acute liver failure may progress to exhibit cerebral dysfunction even HEPATIC COMA depending on the etiology that includes hepatic ISCHEMIA, drug toxicity, malignant infiltration, and viral hepatitis such as post-transfusion HEPATITIS B and HEPATITIS C. Acute Hepatic Failure,Fulminant Hepatic Failure,Fulminating Hepatic Failure,Hepatic Failure, Fulminant,Liver Failure, Fulminant,Acute Liver Failure,Fulminating Liver Failure,Hepatic Failure, Acute,Failure, Acute Hepatic,Failure, Acute Liver,Fulminant Hepatic Failures,Fulminant Liver Failure,Fulminant Liver Failures,Fulminating Hepatic Failures,Fulminating Liver Failures,Hepatic Failure, Fulminating,Liver Failure, Fulminating
D019032 Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization A mass spectrometric technique that is used for the analysis of large biomolecules. Analyte molecules are embedded in an excess matrix of small organic molecules that show a high resonant absorption at the laser wavelength used. The matrix absorbs the laser energy, thus inducing a soft disintegration of the sample-matrix mixture into free (gas phase) matrix and analyte molecules and molecular ions. In general, only molecular ions of the analyte molecules are produced, and almost no fragmentation occurs. This makes the method well suited for molecular weight determinations and mixture analysis. Laser Desorption-Ionization Mass Spectrometry, Matrix-Assisted,MALD-MS,MALDI,Mass Spectrometry, Matrix-Assisted Laser Desorption-Ionization,Mass Spectroscopy, Matrix-Assisted Laser Desorption-Ionization,Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry,Spectroscopy, Mass, Matrix-Assisted Laser Desorption-Ionization,MALDI-MS,MS-MALD,SELDI-TOF-MS,Surface Enhanced Laser Desorption Ionization Mass Spectrometry,Laser Desorption Ionization Mass Spectrometry, Matrix Assisted,MALDI MS,Mass Spectrometry, Matrix Assisted Laser Desorption Ionization,Mass Spectroscopy, Matrix Assisted Laser Desorption Ionization,Matrix Assisted Laser Desorption Ionization Mass Spectrometry

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