Purification and characterization of Atlantic salmon (Salmo salar) fibrinogen.
This study describes a purification protocol of salmon fibrinogen that gives a consumable and highly clottable fibrinogen. Some characteristics of salmon and human fibrinogen are compared. Fibrinogen was purified from barium sulphate adsorbed plasma of Atlantic salmon, using two steps of 25% ammonium sulphate precipitation followed by ultrafiltration. The clottability of the purified salmon fibrinogen was 91%. The Aalpha chains of salmon fibrinogen were heterogeneous with a molecular mass of 90-110 kDa, compared to approximately 67 kDa of human fibrinogen Aalpha chains. The Bbeta and gamma chains of salmon and human fibrinogen had molecular masses of approximately 55 and 50 kDa, respectively. Western blotting revealed that polyclonal rabbit anti-human fibrinogen antibodies had affinity for the gamma chains of salmon fibrinogen, making it possible to study factor XIII activity in purified salmon fibrinogen. Cross-linking of either gamma-gamma or gamma-alpha chains was not detected upon incubation of the purified fibrinogen with thrombin and calcium alone, but was detected when clotting was performed in plasma indicating absence of factor XIII activity in the purified product.
Manseth
Even
E
Department of Chemistry, Biotechnology and Food Science, Agricultural University of Norway, Chr. M. Falsensgate 1, P.O. Box 5040, N-1432 As, Norway. evert.manseth@ikbm.nlh.no
Skjervold
Per O
PO
Fjaera
Svein O
SO
Brosstad
Frank R
FR
Bjørnson
Stine
S
Flengsrud
Ragnar
R
D000818:Animals; D001780:Blood Coagulation Tests; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D003429:Cross Reactions; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D011817:Rabbits; D020317:Salmo salar; D013917:Thrombin / Q000378:metabolism
C464794:fibrinogen Aalpha; D005340:Fibrinogen; D013917:Thrombin; D002118:Calcium