Binding properties on Sepharose insolubilized fibrinogen and fibrin, of various species of fibrinogen and fibrin solubilized in plasma.
Radiolabelled tracers of fibrinogen, fibrin des-AA and fibrin des-AABB were solubilized in recalcified, prothrombin depleted plasma, adding either 125I-fibrin des-AA or 125I-fibrin des-AABB together with 131I-fibrinogen, and subsequently subjected to affinity chromatography, utilizing short columns of Sepharose insolubilized preparations of fibrinogen, fibrin des-AA and fibrin des-AABB, respectively. Two naturally occurring fibrinogen species, of high molecular weight (HMW; m.w. 340.000) and of low molecular weight (LMW; m.w. 305.000) exhibited similar binding characteristics, as judged by adsorption and desorption experiments. In subsequent studies all tracer preparations were derived from HMW-fibrinogen. Sepharose insolubilized fibrinogen favoured the adsorption of soluble fibrins as compared to fibrinogen in solution; the adsorption of soluble des-AA fibrin was similar to that of soluble des-AABB fibrin. To insolubilized fibrin, adsorption of soluble tracers of fibrinogen and fibrins increased considerably, and soluble fibrins were no longer preferentially adsorbed. The latter observation was supported by similar desorption characteristics of these tracers. These findings may indicate that the E-domains of soluble fibrin become largely inaccessible to the D-domains of Sepharose insolubilized fibrinogen, probably due to complexing fibrinogen in plasma. Furthermore, adsorption was largely related to the a-epitope of insolubilized fibrin.
D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D005337:Fibrin / Q000302:isolation & purification / Q000494:pharmacology*; D005338:Fibrin Fibrinogen Degradation Products / Q000302:isolation & purification / Q000494:pharmacology; D005340:Fibrinogen / Q000302:isolation & purification / Q000494:pharmacology*; D007457:Iodine Radioisotopes; D011485:Protein Binding; D012685:Sepharose / Q000494:pharmacology*; D012995:Solubility
D005338:Fibrin Fibrinogen Degradation Products; D007457:Iodine Radioisotopes; C011468:fibrinmonomer; D005337:Fibrin; D005340:Fibrinogen; D012685:Sepharose