Ultra-rapid preparation of milligram quantities of the purified polypeptide chains of human fibrinogen.
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.
Raut
S
S
Division of Haematology, National Institute for Biological Standards and Control, Potters Bar, Hertfordshire, UK.
Corran
P H
PH
Gaffney
P J
PJ
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
D010455:Peptides; D005340:Fibrinogen