Use of Affinity Chromatography for Determination of Dissociation Constants of Complexes of Trypsin and Chymotrypsin with their Free and Immobilized Inhibitors

1978 ◽  
pp. 451-452 ◽  
Author(s):  
Jaroslava Turkova
Keyword(s):  
Affinity Chromatography ◽  
Dissociation Constants

Quantitative Assessment of Soluble Fibrin in Plasma by Affinity Chromatography – A Comparative Study with desAA-Fibrin, desAABB-Fibrin and Fibrinogen

1985 ◽  
Vol 54 (02) ◽  
pp. 533-538 ◽  
Author(s):  
Wilfried Thiel ◽  
Ulrich Delvos ◽  
Gert Müller-Berghaus
Keyword(s):  
Affinity Chromatography ◽  
Calcium Ions ◽  
Quantitative Assessment ◽  
Fluid Phase ◽  
Soluble Fibrin ◽  
Binding Characteristics ◽  
Different Temperatures ◽  
Immobilized Proteins ◽  
Sepharose 4B

SummaryA quantitative determination of soluble fibrin in plasma was carried out by affinity chromatography. For this purpose, desAA-fibrin and fibrinogen immobilized on Sepharose 4B were used at the stationary side whereas batroxobin-induced 125I-desAA-fibrin or thrombin-induced 125I-desAABB-fibrin mixed with plasma containing 131I-fibrinogen represented the fluid phase. The binding characteristics of these mixtures to the immobilized proteins were compared at 20° C and 37° C. Complete binding of both types of fibrin to the immobilized desAA-fibrin was always seen at 20° C as well as at 37° C. However, binding of soluble fibrin was accompanied by substantial binding of fibrinogen that was more pronounced at 20° C. Striking differences depending on the temperature at which the affinity chromatography was carried out, were documented for the fibrinogen-fibrin interaction. At 20° C more than 90% of the applied desAA-fibrin was bound to the immobilized fibrinogen whereas at 37° C only a mean of 17% were retained at the fibrinogen-Sepharose column. An opposite finding with regard to the tested temperature was made with the desAABB-fibrin. Nearly complete binding to insolubilized fibrinogen was found at 37° C (95%) but only 58% of the desAABB-fibrin were bound at 20° C. The binding patterns did not change when the experiments were performed in the presence of calcium ions. The opposite behaviour of the two types of soluble fibrin to immobilized fibrinogen at the different temperatures, together with the substantial binding of fibrinogen in the presence of soluble fibrin to insolubilized fibrin in every setting tested, devaluates affinity chromatography as a tool in the quantitative assessment of soluble fibrin in patients’ plasma.

Factors Influencing the Separation of Glu-Plasminogen Affinity Forms I and II by Affinity Chromatography

1984 ◽  
Vol 52 (03) ◽  
pp. 347-349 ◽  
Author(s):  
Daan W Traas ◽  
Bep Hoegee-de Nobel ◽  
Willem Nieuwenhuizen
Keyword(s):  
Affinity Chromatography ◽  
Dissociation Constants ◽  
Ionic Composition ◽  
Chloride Ions ◽  
Factors Influencing ◽  
Two Factors ◽  
Human Plasminogen

SummaryNative human plasminogen, the proenzyme of plasmin (E. C. 3.4.21.7) occurs in blood in two well defined forms, affinity forms I and II. In this paper, the feasibility of separating these forms of human native plasminogen by affinity chromatography, is shown to be dependent on two factors: 1) the ionic composition of the buffer containing the displacing agent: buffers of varying contents of sodium, Tris, phosphate and chloride ions were compared, and 2) the type of adsorbent. Two adsorbents were compared: Sepharose-lysine and Sepharose-bisoxirane-lysine. Only in the phosphate containing buffers, irrespective of the type of adsorbent, the affinity forms can be separated. The influence of the adsorbent can be accounted for by a large difference in dissociation constants of the complex between plasminogen and the immobilized lysine.

Determination of acidic dissociation constants of L-phenylalanyl-glycine and L-alanyl-L-alanine in water using ab initio methods

2014 ◽  
Vol 2 (42) ◽  
pp. 263-263
Author(s):  
Farhoush Kiani ◽  
Mahmoud Tajbakhsh ◽  
Fereydoon Ashrafi ◽  
Nesa Shafiei ◽  
Azar Bahadori ◽  
...  
Keyword(s):  
Ab Initio ◽  
Dissociation Constants ◽  
Ab Initio Methods ◽  
Acidic Dissociation

Determination of glycated hemoglobin by affinity chromatography

1987 ◽  
Vol 168 (1) ◽  
pp. 81-86 ◽  
Author(s):  
Gerhard Oremek ◽  
Ulrich B. Seiffert ◽  
Günter Schmid
Keyword(s):  
Affinity Chromatography ◽  
Glycated Hemoglobin

scholarly journals Determination of equilibrium dissociation constants for recombinant antibodies by high-throughput affinity electrophoresis

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Yuchen Pan ◽  
Eric K. Sackmann ◽  
Karolina Wypisniak ◽  
Michael Hornsby ◽  
Sammy S. Datwani ◽  
...  
Keyword(s):  
High Throughput ◽  
Dissociation Constants ◽  
Recombinant Antibodies ◽  
Affinity Electrophoresis ◽  
Equilibrium Dissociation Constants ◽  
Equilibrium Dissociation

scholarly journals Circular dichroism determination of class I MHC-peptide equilibrium dissociation constants

1997 ◽  
Vol 6 (8) ◽  
pp. 1771-1773 ◽  
Author(s):  
Chantal S. Morgan ◽  
James M. Holton ◽  
Barry D. Olafson ◽  
Pamela J. Bjorkman ◽  
Stephen L. Mayo
Keyword(s):  
Circular Dichroism ◽  
Dissociation Constants ◽  
Class I ◽  
Class I Mhc ◽  
Equilibrium Dissociation Constants ◽  
Equilibrium Dissociation ◽  
Circular Dichroïsm
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