title: Affinity purification of plasma proteins: characterization of six affinity matrices and their application for the isolation of human factor VIII.
abstract: For the purification of coagulation factor VIII, (1,1'-carbonyl-diimidazole [CDI]-activated) Sepharose CL-4B was functionalized with two aminoalkyl and four aminoalkyl-carbamylalkyl ligand-spacer combinations. The affinity matrices were contacted with human plasma. All affinity matrices showed complete adsorption of factor VIII (greater than 90%) and three aminoalkyl-carbamylalkyl Sepharoses gave factor-VIII recoveries of 50-65% and a factor-VIII preparation with a specific activity of 1-2 U factor VIII/mg of protein. Furthermore, no fibrinogen, immunoglobulin G and albumin could be detected in the isolated factor VIII. Optimal results were obtained using the di-methyl-aminopropyl-carbamyl-pentyl-Sepharose affinity matrix.
authors.affiliation: Dept. of Development and Quality Assurance, Netherlands Red Cross Blood Transfusion Service, Amsterdam.
authors.forename: M P
authors.initials: MP
authors.lastname: te Booy
authors.forename: W
authors.initials: W
authors.lastname: Riethorst
authors.forename: A
authors.initials: A
authors.lastname: Faber
authors.forename: J
authors.initials: J
authors.lastname: Over
authors.forename: B W
authors.initials: BW
authors.lastname: König
mesh_terms: D000327:Adsorption; D005169:Factor VIII / Q000302:isolation & purification*; D005340:Fibrinogen / Q000032:analysis; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D012685:Sepharose / Q000031:analogs & derivatives; D012709:Serum Albumin / Q000032:analysis; D014841:von Willebrand Factor / Q000302:isolation & purification
chemical_list: D007074:Immunoglobulin G; D012709:Serum Albumin; D014841:von Willebrand Factor; D005169:Factor VIII; D005340:Fibrinogen; D012685:Sepharose