title: Ultra-rapid preparation of milligram quantities of the purified polypeptide chains of human fibrinogen.
abstract: In order to study the epitopes in fibrin towards which monoclonal antibodies are directed we needed the pure individual polypeptide chains of human fibrinogen in reasonable quantity. We report here a simplified, rapid method of separation of high-purity human fibrinogen chains. Following reduction and S-carboxymethylation of human fibrinogen, the sample was injected directly onto a column of the polymeric reversed-phase perfusion packing POROS 20-R2, and the chains were completely resolved in less than 3 min at a flow-rate of 10 ml/min. The capacity was equivalent to that of a similar sized conventional silica-based column. However the throughput was approximately five to ten times as high. The column was durable and robust in day-to-day use.
authors.affiliation: Division of Haematology, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, UK.
authors.forename: S
authors.initials: S
authors.lastname: Raut
authors.forename: P H
authors.initials: PH
authors.lastname: Corran
authors.forename: P J
authors.initials: PJ
authors.lastname: Gaffney
mesh_terms: D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000302:isolation & purification*; D006801:Humans; D010084:Oxidation-Reduction; D010455:Peptides / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D010455:Peptides; D005340:Fibrinogen