DETERMINATION OF ANTITRYPTIC ACTIVITY IN PROTEINS FROM PEANUT PRODUCTS ISOLATED BY AFFINITY CHROMATOGRAPHY

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.

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Determination of glycated hemoglobin by affinity chromatography

Clinica Chimica Acta ◽

10.1016/0009-8981(87)90269-5 ◽

1987 ◽

Vol 168 (1) ◽

pp. 81-86 ◽

Cited By ~ 9

Author(s):

Gerhard Oremek ◽

Ulrich B. Seiffert ◽

Günter Schmid

Keyword(s):

Affinity Chromatography ◽

Glycated Hemoglobin

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

Enzyme Engineering ◽

10.1007/978-1-4684-6985-1_93 ◽

1978 ◽

pp. 451-452 ◽

Cited By ~ 2

Author(s):

Jaroslava Turkova

Keyword(s):

Affinity Chromatography ◽

Dissociation Constants

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Determination of the carbohydrate-binding site of Bauhinia purpurea lectin by affinity chromatography

Journal of Chromatography A ◽

10.1016/0021-9673(92)80114-a ◽

1992 ◽

Vol 597 (1-2) ◽

pp. 221-230 ◽

Cited By ~ 6

Author(s):

Kazuo Yamamoto ◽

Yukiko Konami ◽

Toshiaki Osawa

Keyword(s):

Affinity Chromatography ◽

Binding Site ◽

Carbohydrate Binding ◽

Bauhinia Purpurea ◽

Bauhinia Purpurea Lectin ◽

Carbohydrate Binding Site

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Determination of the Kinetic Rate Constant of Cyclodextrin Supramolecular Systems by High-Performance Affinity Chromatography

Methods in Molecular Biology - Affinity Chromatography ◽

10.1007/978-1-4939-2447-9_24 ◽

2015 ◽

pp. 309-319

Author(s):

Jiwen Zhang ◽

Haiyan Li ◽

Lixin Sun ◽

Caifen Wang

Keyword(s):

Affinity Chromatography ◽

Rate Constant ◽

High Performance ◽

Supramolecular Systems ◽

Kinetic Rate Constant ◽

Kinetic Rate ◽

High Performance Affinity Chromatography

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Solid-phase radioimmunoassay for morphine, with use of an affinity-purified morphine antibody.

Clinical Chemistry ◽

10.1093/clinchem/24.2.339 ◽

1978 ◽

Vol 24 (2) ◽

pp. 339-342 ◽

Cited By ~ 4

Author(s):

M Steiner ◽

J L Spratt

Keyword(s):

Affinity Chromatography ◽

Correlation Coefficient ◽

Solid Phase ◽

Rapid Determination ◽

Biological Fluids ◽

Inhibitory Effect ◽

Major Metabolite ◽

Solid Phase Radioimmunoassay ◽

Related Compounds

Abstract Morphine antibody purified by affinity chromatography was used to develop a solid-phase radioimmunoassay for morphine in polystyrene tubes. The tubes are coated with an appropriate concentration of the purified antibody, rinsed three times with buffered saline, and stored at -15 degrees C. Using tritiated dihydromorphine, we determined competitive morphine binding by difference when the radioactivity in the assay supernates was measured after incubation (1 h, 37 degrees C). Five standard curves, with use of serum equivalents of morphine ranging from 0 to 6 mug/liter, were linear and had a mean correlation coefficient of 0.98. Uncer conditions of the assay, levorphanol was comparable to morphine in its inhibitory effect on binding of labeled dihydromorphine, whereas dextrorphan was essentially inactive. Morphine-3-glucuronide, a major metabolite, is 55-fold less inhibitory in terms of its capacity to displace the radiolabel. We believe that the sensitivity of the technique, coupled with the simplicity of nonseparatory sampling, renders the system suitable for rapid determination of morphine and related compounds in biological fluids.

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