scholarly journals Studies on the mechanism of LDH-IgG complex formation using 5'-AMP and Cibacron Blue F3G-A affinity chromatography

1988 ◽  
Vol 32 (4) ◽  
pp. 187-195
Author(s):  
Kiyotaka Fujita ◽  
Ikunosuke Sakurabayashi ◽  
Tadashi Kawai ◽  
Mutsuko Kusanagi ◽  
Yoshimi Teramura
Keyword(s):  
Complex Formation ◽  
Affinity Chromatography ◽  
Cibacron Blue

scholarly journals Complex Formation of Covalently Crosslinked Fibrin Oligomers with Agarose Coupled Fibrinogen and Fibrin

1977 ◽  
Author(s):  
R. von Hugo ◽  
R. Hafter ◽  
A. Stemberger ◽  
H. Graeff
Keyword(s):  
Molecular Weight ◽  
Complex Formation ◽  
Affinity Chromatography ◽  
High Molecular Weight ◽  
Calcium Chloride ◽  
Gel Filtration ◽  
Void Volume ◽  
Soluble Fibrin ◽  
Derivatives Of

Crosslinked high molecular weight derivatives of fibrin (fibrinoligomers) were observed during intravascular coagulation. It was the purpose of this study to investigate the complex formation of fibrin oligomers with fibrinogen and fibrinmonomer. Fibrinogen coupled to agarose (Fg-ag) as well as fi-brinmonomer coupled to agarose (Fm-ag) was used as substrate. Soluble oligomers of fibrin were produced by incubating fibrinogen with thrombin, calcium-chloride, cystein and F XIII. They were separated from fibrinogen by gel filtration. Γ-dimers were demonstrated in fractions from the void volume and the shoulder prior to the fibrinogen peak. These fractions were subjected to affinity chromatography. Crosslinked oligomers of fibrin were not adsorbed on Fg-ag, yet adsorption occured on Fm-ag. This indicates that fibrin oligomers have no affinity to fibrinogen, yet readily form complexes with fibrin. This could mean that in vivo they compete with fibrinogen for the fibrinmonomer part of soluble fibrin monomer complexes, and hence have a tendency to increase in size.

A lactate dehydrogenase-immunoglobulin G1 complex, not blocked by anti-idiotype antibody, in a patient with IgG1-lambda type M-proteinemia.

1987 ◽  
Vol 33 (8) ◽  
pp. 1478-1483 ◽  
Author(s):  
K Fujita ◽  
I Sakurabayashi ◽  
M Kusanagi ◽  
T Kawai
Keyword(s):  
Lactate Dehydrogenase ◽  
Complex Formation ◽  
Affinity Chromatography ◽  
Binding Site ◽  
Binding Capacity ◽  
Isoenzyme Pattern ◽  
M Protein ◽  
Light Chains ◽  
Ldh Isoenzymes ◽  
Sepharose 4B

Abstract The serum of a patient with IgG1-lambda type M-proteinemia showed an abnormal isoenzyme pattern for lactate dehydrogenase (LDH, EC 1.1.1.27). By affinity chromatography, we showed that four isoenzymes (LDH2, LDH3, LDH4, and LDH5) were bound to the M-protein. This complex formation was not blocked by anti-idiotype antibody, even though the binding capacity of IgG was exclusively located in the Fab region of the molecule. Moreover, heavy and light chains of the patient's IgG, obtained by reduction, separately had affinities for each of the LDH isoenzymes. LDH-IgG complex was easily dissociated by affinity chromatography on 5'-AMP-Sepharose 4B or by added NADH. We propose the following hypothesis for the LDH-IgG complex formation: LDH can recognize the gamma-Fab region of IgG at the NAD+ binding site of the molecule, but the affinity of the LDH molecule for immunoglobulin is much weaker than that for NADH or 5'-AMP.

Affinity chromatography of human plasma gelsolin with polyphosphate compounds on immobilized Cibacron Blue F3GA

1990 ◽  
Vol 526 ◽  
pp. 397-406 ◽  
Author(s):  
Hiroaki Ito ◽  
Hideo Yamamoto ◽  
Yoshihiro Kimura ◽  
Hiroshi Kambe ◽  
Toshikazu Okochi ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Human Plasma ◽  
Cibacron Blue ◽  
Plasma Gelsolin ◽  
Cibacron Blue F3ga

Separation of Fibrinogen Degradation Products (FDP) by Means of Adsorption to Insolubilized Fibrinmonomer (FM-ag)

1979 ◽  
Vol 41 (04) ◽  
pp. 677-686 ◽  
Author(s):  
D L Heene ◽  
F R Matthias ◽  
Z Wegrzynowicz ◽  
G Hocke
Keyword(s):  
Complex Formation ◽  
Affinity Chromatography ◽  
Degradation Products ◽  
Preparative Method ◽  
Disc Electrophoresis ◽  
Essential Difference ◽  
Fibrinogen Degradation Products ◽  
Adsorption Characteristics

SummaryThe phenomenon of complex formation between fibrinmonomer and fibrinogen degradation products was investigated by means of adsorption of FDP to insolubilized thrombin- modified fibrinogen (FM-ag). Since it could be demonstrated that there are different adsorption characteristics for early FDP and late FDP, the possibility of separation of FDP by means of affinity chromatography on FM-ag columns was evaluated using plasmic digests of 3H-Ac-labelled fibrinogen. The identification of FDP was performed by disc-electrophoresis. The results indicate that the adsorption of early FDP is comparable to the behaviour of fibrinogen, whereas late FDP show essential difference in the affinity towards FM-ag, evident by the result that fragment E adsorbs only to a minimal extent. Fragments D and E derived from fibrinogen as well as from non-crosslinked fibrin, revealed identical adsorption characteristics. Under specified conditions the procedure is suitable as a preparative method for the separation of fragments D and E.

Circulating IgG-LD complex, dissociable by addition of NAD+.

1982 ◽  
Vol 28 (1) ◽  
pp. 236-239 ◽  
Author(s):  
F Gorus ◽  
W Aelbrecht ◽  
B Van Camp
Keyword(s):  
Lactate Dehydrogenase ◽  
Autoimmune Disease ◽  
Complex Formation ◽  
Affinity Chromatography ◽  
Conformational Changes ◽  
Active Site ◽  
Nicotinamide Adenine Dinucleotide ◽  
Binding Sites ◽  
Gel Filtration ◽  
Binding Studies

Abstract Macromolecular LD (lactate dehydrogenase, EC 1.1.1.27) was present in the serum of a patient suffering from idiopathic fibrosis of the lung and presenting signs of autoimmune disease. By using gel filtration and affinity chromatography techniques, the vast majority of the patient's serum LD activity was shown to consist of LD-IgG complexes, which dissociated in the presence of added nicotinamide adenine dinucleotide (NAD+). Binding studies with tritiated NAD+ indicated that complex formation was not ascribable to a lack of circulating cofactor. The most likely explanation for the complex formation was the existence of LD binding sites on IgG molecules. The disruption of the complex by NAD+ might be explained by a competition between IgG molecules and NAD+ for the LD active site or by conformational changes induced in the LD molecules on binding of NAD+.

Effect of albumin on binding and recovery of enzymes in affinity chromatography on Cibacron Blue

1983 ◽  
Vol 130 (2) ◽  
pp. 481-484 ◽  
Author(s):  
Candadai S. Ramadoss ◽  
Janusz Steczko ◽  
John W. Uhlig ◽  
Bernard Axelrod
Keyword(s):  
Affinity Chromatography ◽  
Cibacron Blue
Sign in / Sign up

Export Citation Format

Share Document