Temperature dependent dissociation of soluble fibrin monomer complexes demonstrated by agarose gel filtration

1980 ◽  
Vol 20 (3) ◽  
pp. 325-333 ◽  
Author(s):  
R. Hafter ◽  
R. von Hugo ◽  
M. Baumgärtner ◽  
F.K. Hiller ◽  
H. Graeff
Keyword(s):  
Gel Filtration ◽  
Agarose Gel ◽  
Temperature Dependent ◽  
Soluble Fibrin ◽  
Fibrin Monomer

Soluble Fibrin Monomer Complexes Demonstrated by Agarose Gel Filtration and by Adsorption on Insolubilized Fibrinogen

1975 ◽  
Vol 34 (01) ◽  
pp. 216-222 ◽  
Author(s):  
R von Hugo ◽  
R Hafter ◽  
A Stemberger ◽  
H Graeff
Keyword(s):  
Molecular Weight ◽  
Electrophoretic Mobility ◽  
Gel Filtration ◽  
Adsorption Chromatography ◽  
Agarose Gel ◽  
Soluble Fibrin ◽  
Relative Electrophoretic Mobility ◽  
Fibrin Monomer

SummaryIncubation of fibrinogen with small amounts of thrombin resulted in the occurrence of soluble fibrin monomer complexes. These complexes consisted predominantly of a derivative with a higher molecular weight than that of fibrinogen. It was characterized by its relative electrophoretic mobility in 5% PAA gel (0.28 × 10-5 cm2/V × sec) and its elution position prior to the fibrinogen peak following gel filtration. Using adsorption chromatography on insolubilized fibrinogen the derivative dissociated at a ratio of almost 1 : 1 into one part which was adsorbed and into fibrinogen which was not adsorbed. The part which was adsorbed seemed to be the thrombin mediated fibrin monomer. This study confirms the concept that dissociable dimeric fibrinogen-fibrin monomer complexes occur after limited action of thrombin on fibrinogen.

Soluble Fibrin Monomer Complexes Demonstrated by Agarose Gel Filtration and by Adsorption on Insolubilized Fibrinogen. A Comparative Study

1975 ◽  
Author(s):  
von R. Hugo ◽  
R. Hafter ◽  
A. Stemberger ◽  
H. Graeft
Keyword(s):  
Molecular Weight ◽  
Comparative Study ◽  
Electrophoretic Mobility ◽  
Gel Filtration ◽  
Adsorption Chromatography ◽  
Agarose Gel ◽  
Soluble Fibrin ◽  
Relative Electrophoretic Mobility ◽  
Fibrin Monomer

Incubation of fibrinogen with small amounts of thrombin resulted in the occurrence of soluble fibrin monomer complexes. These complexes consisted predominantly of a derivative with a higher molecular weight than that of fibrinogen. It was characterized by its relative electrophoretic mobility in 5% PAA gel (0.28 × 10-5 cm2/V×sec) and its elution position prior to the fibrinogen peak following gel filtration. Using adsorption chromatography on insolubilized fibrinogen the derivative dissociated at a ratio of almost 1 : 1 into one part which was adsorbed and into fibrinogen which was not adsorbed. The part which was adsorbed seemed to be the thrombin mediated fibrin monomer. This study confirms the concept that dissociable dimeric fibrinogonfibrin monomer complexes occur after limited action of thrombin on fibrinogen.(Supported by DFG ; SFB 51, grant no. 2/Gra – B/6.)

Studies on Soluble Fibrin Complexes as a Function of pH

1975 ◽  
Author(s):  
Y. Benabid ◽  
E. Concord ◽  
M. Suscillon
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Gel Chromatography ◽  
Agarose Gel ◽  
Monomer Mixture ◽  
Soluble Fibrin ◽  
Molecular Weights ◽  
Fibrin Monomer

Purified fibrinogen solutions, incubated with thrombin. CNBr. Sepharose, were subjected to agarose gel chromatography and eluted at different pH (6.5; 7.5; 8.5). Among high molecular weight derivatives formed by thrombin, the major component was a dimer. Gel chromatography at pH 8.5 showed a complexes peak distinct of that from fibrinogen, whereas at pH 6.5, only the fibrinogen peak appeared: fibrin monomer was eluted with fibrinogen as demonstrated by polyacrylamid gel electrophoresis 3.75% pH 8.9. SDS urea electrophoresis after reduction indicated that complexes peak contained two α-chains (α and α′). When fibrinogen was incubated with thrombin in the presence of FSF and calcium, several derivatives with higher and higher molecular weights were formed besides the dimer, and elution profiles of chromatography were identical at pH 6.5 and 8.5, thus indicating stable complexes formation. If fibrinogen-fibrin monomer mixture was subjected to FSF action at different pH, no complexes were formed at pH 6.5. These results confirm that at pH 6.5, any association was prevented.

scholarly journals Soluble Fibrin Complexes During Hormonal Contraception

1977 ◽  
Author(s):  
F. Asbeck ◽  
J. Bebber ◽  
J. van de Loo
Keyword(s):  
Oral Contraceptive ◽  
Degradation Products ◽  
Gel Filtration ◽  
Low Grade ◽  
Agarose Gel ◽  
Fibrinogen Degradation Products ◽  
Soluble Fibrin ◽  
Healthy Women ◽  
Derivatives Of ◽  
Precipitable Protein

In a prospective study, 2o healthy women (16–35 yrs.) were treated with an oral contraceptive (o,25 mg D-norgestrel, o,o5 mg ethinyl-estradiol). Derivatives of fibrinogen and fibrin were investigated before the beginning of medication, after the 1st cycle, and after the 3rd cycle. The investigations included: determination of thrombin-clottable protein, cold precipitable protein (4°C) , ethanol precipitable protein (11,5%), ethanol gelation test, protamin sulfate test, fibrin-fibrinogen degradation products in serum, and quantitation of high-and low molecular weight derivatives of fibrinogen and fibrin by agarose gel filtration of the fresh patient plasmas.There was a significant increase of thrombin clottable protein (212 vs. 236 mg/dl, p<0,001) and an increase of ethanol precipitable protein (117 vs. 168 mg/dl, p<0,003). There was no change in the cold precipitable protein nor in the paracoagulation phenomena. Fibrin-fibrinogen degradation products were not elevated. By agarose gel filtration, a significant increase in soluble fibrin complexes (p<0,02) could be demonstrated; the highest concentration of the SFC elu-ted in the position of the dimers.The study confirms a low grade activation of the coagulation system even in healthy women taking a low dose estrogen containing oral contraceptive.

In Vitro Formation of Fibrin-Monomer Complexes in the Presence of Isotope Labelled Fibrinogen-Fibrin Degradation Products. An Agarose Gel Filtration Study

1975 ◽  
Author(s):  
F. Asbeck ◽  
van de J. Loo
Keyword(s):  
Molecular Weight ◽  
Complex Formation ◽  
Degradation Products ◽  
Gel Filtration ◽  
Agarose Gel ◽  
Fibrinogen Degradation Products ◽  
Molecular Weights ◽  
Fibrin Degradation Products ◽  
Fibrin Monomer

Human citrated plasmas were mixed with purified 131I-fibrinogen and 131I-fibrinogen degradation products (FDP) or 125I-fibrin degradation products (fdp). After incubation with small amounts of thrombin (0.01–0.02 imits/ml Pl.), these mixtures were gel filtrated on Biogel A5m columns and the elution patterns of the 131I- and -labelled materials were determined.In control experiments without thrombin incubation, no complex formation between fibrinogen, FDP or fdp in citrated plasmas could be detected. This was even true for fdp with a higher molecular weight than fibrinogen.After thrombin incubation, up to 11% fibrin-monomer complexes were formed. Irrespective of their molecular weights, labelled fdp were not incorporated into these complexes.Only large FDP – presumably derivative X – did partially copolymerize with fibrin-monomer complexes in citrated plasmas.

scholarly journals Analysis of Soluble Fibrin Complexes in Patients with Intra Vascular Coagulation

1977 ◽  
Author(s):  
W. Edgar ◽  
C. McKillop ◽  
P. W. Howie ◽  
C. D. Forbes ◽  
C. R. M. Prentice
Keyword(s):  
Major Part ◽  
Polypeptide Chain ◽  
Gel Filtration ◽  
Chain Structure ◽  
Agarose Gel ◽  
Soluble Fibrin ◽  
Α Chain ◽  
Sepharose 4B ◽  
Soluble Complexes

We have analysed the polypeptide chain structure of soluble fibrin complexes in patients with pre-eclampsia and during defibrination with ancrod (Arvin). In pre-eclampsia soluble fibrin measured by fibrinogen-sepharose 4B chromatography was increased in comparison with normal pregnant women. The isolated fibrin contained intact α, β and δ chains but no crosslihked γ chains. A correlation (r = 0.959 P <0.001) was found between the concentration of soluble fibrin and soluble complexes as measured by agarose gel filtration, suggesting these complexes consist mainly of fibrin-fibrinogen dimers. In vitro, the binding of soluble fibrin to fibrinogen-sepharose depended on the structure of the complexes and temperature. At 37°C intact fibrin, prepared with ancrod, was bound to the column, but fibrin lacking intact α chain could be eluted at 37°C. The soluble fibrin complexes in patients having therapy with ancrod could be separated into two;the greater part had reduced α chain and were eluted from the column at 3 7°C, whereas the smaller part had intact α chain and remained bound at 37°C. Bio-Gel A5m chromatography carried out at 37°C and 20°C also indicated that the major part of soluble complexes produced during ancrod defibrination were unstable at 37°C.

Detection of Soluble Fibrin Monomer Complexes in Blood by Means of Protamine Sulphate Test

1968 ◽  
Vol 20 (01/02) ◽  
pp. 044-049 ◽  
Author(s):  
B Lipiński ◽  
K Worowski
Keyword(s):  
Human Subjects ◽  
Coronary Thrombosis ◽  
Mean Value ◽  
Healthy Human ◽  
Protamine Sulphate ◽  
Soluble Fibrin ◽  
Fibrin Monomer ◽  
Dog Plasma ◽  
The Mean ◽  
Precipitable Protein

SummaryIn the present paper described is a simple test for detecting soluble fibrin monomer complexes (SFMC) in blood. The test consists in mixing 1% protamine sulphate with diluted oxalated plasma or serum and reading the optical density at 6190 Å. In experiments with dog plasma, enriched with soluble fibrin complexes, it was shown that OD read in PS test is proportional to the amount of fibrin recovered from the precipitate. It was found that SFMC level in plasma increases in rabbits infused intravenously with thrombin and decreases after injection of plasmin with streptokinase. In both cases PS precipitable protein in serum is elevated indicating enhanced fibrinolysis. In healthy human subjects the mean value of OD readings in plasma and sera were found to be 0.30 and 0.11, while in patients with coronary thrombosis they are 0.64 and 0.05 respectively. The origin of SFMC in circulation under physiological and pathological conditions is discussed.

Soluble Fibrin Complexes and Fibrinogen Heterogeneity in Diabetes mellitus

1980 ◽  
Vol 44 (03) ◽  
pp. 130-134 ◽  
Author(s):  
E B Tsianos ◽  
N E Stathakis
Keyword(s):  
Diabetes Mellitus ◽  
Molecular Weight ◽  
Plasma Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Lower Molecular Weight ◽  
Polyacrylamide Gels ◽  
Soluble Fibrin ◽  
Α2 Macroglobulin ◽  
Patients With Diabetes

SummaryThe presence of soluble fibrin complexes (SFC) measured by gel filtration of plasma on 4% agarose columns, fibrinogen heterogeneity on 3.5% SDS-polyacrylamide gels and the concentrations of several plasma proteins were evaluated in 39 patients with diabetes mellitus (DM) and 19 matched control subjects. A small but significant increase of SFC was found in DM (p<0.01). On individual basis 51.2% of the patients had increased SFC (>M + 2 SD of the controls). Polyacrylamide gel electrophoresis of the SFC showed no evidence of cross-linking or proteolysis. Plasma clots formed in the presence of EDTA and trasylol were analysed in SDS-polyacrylamide gels in a normal and two lower molecular weight fibrin bands (band I, II, III). The percentage of band I fibrinogen was in diabetics (65.3 ± 4.7%) lower than that of the controls (71.8 ± 4.5%) (p < 0.01). Fibrinogen levels, antithrombin III, α1-antitrypsin, α2-macroglobulin and plasminogen were significantly increased in DM. We suggest that in DM there is an enhancement of intravascular fibrin formation and accelerated fibrinogen degradation to lower molecular weight forms.

Identification of Fibrinogen Derivatives in Plasma Samples

1972 ◽  
Vol 27 (03) ◽  
pp. 610-618 ◽  
Author(s):  
H Graeff ◽  
R von Hugo
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Gel Chromatography ◽  
Agarose Gel ◽  
Plasma Samples ◽  
Methodological Study ◽  
Parent Molecule ◽  
Electrophoretic Mobilities ◽  
Fibrin Monomer

SummaryThe observation of fibrinogen derivatives with a molecular weight higher than the parent molecule in human cases of DIC initiated the present methodological study. These derivatives were identified by the following methods : 2.5 M β-alanine precipitation of the plasma samples, PAA gel electrophoresis, intra gel immunoprecipitation and agarose gel chromatography. In the plasma of a patient with severe eclampsia and laboratory signs of DIC two derivatives with a molecular weight higher than that of fibrinogen were identified according to their relative electrophoretic mobilities: 0.18 and 0.28 × 10−5 cm2/V × sec (fibrinogen: 0.43 × 10−5 cm2/V × sec). Electrophoretic studies in the presence of 5 M urea indicated that the 0.28 derivative is a complex probably formed by fibrinogen and a fibrin monomer.

Sign in / Sign up

Export Citation Format

Share Document