Isolation of fibrinogen A alpha-chain by affinity chromatography on concanavalin A-sepharose.
A method was developed for isolation of the A alpha-chain from S-carboxamidomethylated fibrinogen. A mixture of the three constituent polypeptide chains of human fibrinogen was applied onto a column of concanavalin A-Sepharose. While the carbohydrate-free A alpha-chain was not delayed on the affinity chromatography column, both glycosylated subunit chains, B beta- and gamma-chain, were adsorbed to the insolubilized lectin and were quantitatively eluted from the column with 0.2 M methyl-alpha-D-mannoside. SDS-electrophoresis on polyacrylamide gel was employed for analysis of chromatographic fractions. Complete recovery of the A alpha-chain was observed. The described procedure is very simple and permits isolation of large amounts of pure A alpha-chain from S-carboxyamidomethylated fibrinogen.
Bögli
C
C
Central Hematology Laboratory, Inselspital, Bern, Switzerland.
Hofer
A
A
Furlan
M
M
D002846:Chromatography, Affinity; D005338:Fibrin Fibrinogen Degradation Products; D005340:Fibrinogen / Q000302:isolation & purification*; D005344:Fibrinopeptide A / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D012685:Sepharose / Q000031:analogs & derivatives
C053898:Aalpha 1-21 peptide; D005338:Fibrin Fibrinogen Degradation Products; C028326:concanavalin A-sepharose; D005344:Fibrinopeptide A; D005340:Fibrinogen; D012685:Sepharose