Preparation and Utilization of a Biosensor Based on Galactose Oxidase

1992 ◽  
Vol 57 (11) ◽  
pp. 2287-2294 ◽  
Author(s):  
Eva Vrbová ◽  
Jitka Pecková ◽  
Miroslav Marek
Keyword(s):  
Oxygen Sensor ◽  
Specific Activity ◽  
Galactose Oxidase ◽  
Collagen Membrane ◽  
Michaelis Constant ◽  
Nylon Mesh ◽  
Native Collagen ◽  
The Stability ◽  
Galactose Content

An enzyme electrode for D-galactose determination was prepared by fixation of a carrier with immobilized galactose oxidase (E.C. 1.1.3.9) or coimmobilized galactose oxidase and catalase (E.C. 1.11.1.6) to a Clark-type oxygen sensor. The enzymes were immobilized either on a partially hydrolyzed nylon mesh or on a native collagen membrane using the Ugi reaction with cyclohexyl isocyanide and glutaraldehyde. The biosensors were characterized by the specific activity of the immobilized galactose oxidase, the apparent Michaelis constant KM(app.), and the stability expressed by time and a number of the performed analyses. The substrate specificity of the biosensor and the effect of pH and temperature of the reaction mixture on the response magnitude were also tested. The prepared biosensor was used for the determination of D-galactose content in samples of blood plasma and serum of patients with suspected galactosemia.

Preparation of enzyme electrode for D-glucose determination by immobilization of glucose oxidase on collagen membrane

1990 ◽  
Vol 55 (10) ◽  
pp. 2568-2574 ◽  
Author(s):  
Eva Vrbová ◽  
Miroslav Marek
Keyword(s):  
Glucose Oxidase ◽  
Oxygen Sensor ◽  
Immobilized Enzyme ◽  
Sensor Response ◽  
Enzyme Electrode ◽  
Collagen Membrane ◽  
Michaelis Constant ◽  
Glucose Determination ◽  
The Stability

An enzyme electrode for the determination of D-glucose was prepared by immobilization of glucose oxidase (EC 1.1.3.4.) on an activated collagen membrane using glutaraldehyde and the Ugi reaction resp. and by subsequent fixation of the membrane to an oxygen sensor of the Clark type. Two different procedures for the modification of the support, the composition of the reaction mixture and the immobilization time were examined. The electrode prepared was tested as regards the effect of pH and temperature on the magnitude of the response. The range of the linear dependence of the sensor response on substrate concentration (1.7 . 10-5 - 2.0 . 10-3), the apparent Michaelis constant of the immobilized enzyme (KM(app.) = 4.16 . 10-3 mol dm-3) and the stability of the biosensor as function of storage mode and number of assays performed with one electrode were determined. In view of the high stability and linear range of the concentration dependence the enzyme electrode is suitable for the determination of D-glucose in samples analyzed in agricultural and food laboratories.

scholarly journals Influence of Radioactive Contamination of the Sr-90 Terrestrial Ecosystems on the Enzymatic Activity of the Soil

2018 ◽  
Vol 3 (3) ◽  
pp. 137
Author(s):  
Lavrentyeva G.V. ◽  
Zaharova V.R. ◽  
Mirzeabasov O.A. ◽  
Synzynys B.I.
Keyword(s):  
Enzymatic Activity ◽  
Radioactive Contamination ◽  
Catalase Activity ◽  
Specific Activity ◽  
Terrestrial Ecosystems ◽  
Threshold Value ◽  
Control Value ◽  
Reliable Model ◽  
The Stability

One of the most promising methods of soil diagnostics is the determination of the parameters of enzymatic activity. Changes in urease, invertase, dehydrogenase and catalase activity of soil subjected to radioactive contamination with radionuclide Sr-90 have been studied. When the specific activity of Sr-90 in the soil varies in the range from the control value to more than 1.5 kBq / kg, the stability of invertase, urease and dehydrogenase is established. At the same time, the catalase activity of the soil is a sensitive indicator to radioactive contamination, which is described by a reliable model with a threshold value, after which the oppression of the indicator is observed.

An Enzyme Linked Immunosorbent Assay for Determination of Tissue Plasminogen Activator Applied to Patients with Thromboembolic Disease

1983 ◽  
Vol 50 (03) ◽  
pp. 740-744 ◽  
Author(s):  
Nils Bergsdorf ◽  
Torbjörn Nilsson ◽  
Per Wallén
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Specific Activity ◽  
Thromboembolic Disease ◽  
Tissue Type ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type Plasminogen Activator ◽  
Tissue Plasminogen ◽  
Normal Human

SummaryUtilizing the immunoglobulin fraction from a goat antiserum against human uterine tissue plasminogen activator, an enzyme- linked immunoassay for tissue-type plasminogen activator in human plasma has been developed. With the new method, the concentration of t-PA in normal human acidified plasma is found to be 4.0 ± 1.8 (SD) ng/ml. It increases to 12 ng/ml after a tomiquet test, and to 14 ng/ml after strenous physical exercise. In a group of patients with idiopathic thromboembolic disease, the resting t-PA concentration was 5 ng/ml and the post-occlusion value 16 ng/ml. Furthermore, the patients also exhibited a normal post-occlusion rise in the concentration of plasmin-α2-antiplasmin complex. However, in 37% of the post-occlusion patient plasmas, virtually no increase in t-PA could be detected by a specific activity assay. The results indicate that the reason for a defective post-occlusion fibrinolytic activity in a majority of cases may be the presence of increased concentrations of a fast-acting specific t-PA inhibitor.

Determination of Specific Activity of Cholera Chemical Vaccine Components using Cell Culture

2020 ◽  
Vol 36 (3) ◽  
pp. 82-89
Author(s):  
O.V. Gromova ◽  
O.S. Durakova ◽  
S.V. Generalov ◽  
L.F. Livanova ◽  
O.A. Volokh
Keyword(s):  
Cell Culture ◽  
Vibrio Cholerae ◽  
Production Control ◽  
Specific Activity ◽  
Methodological Approach ◽  
Toxin Production ◽  
Submerged Cultivation ◽  
Vaccine Production ◽  
Antigenic Activity

Том 36(2020) №3 стр. 82-89; DOI 10.21519/0234-2758-2020-36-3-82-89А.В. Гаева1*, О.В. Громова1, О.С. Дуракова1, С.В. Генералов1, Л.Ф. Ливанова1, О.А. Волох1 Определение специфической активности компонентов холерной химической вакцины с использованием культуры клеток 1ФКУЗ «Российский научно-исследовательский противочумный институт «Микроб»» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека, Саратов 410005 *[email protected] Поступила - 2019-11-26; После доработки - 2020-03-16; Принята к публикации - 2020-05-15 Список литературы Описаны методы определения динамики продукции токсинов штаммом Vibrio cholerae 569B при глубинном культивировании в биореакторе и антигенной активности специфической фракции холерогена-анатоксина по анатоксинсвязыванию с использованием клеточных культур. Показана высокая степень соответствия результатов, полученных методами, применяемыми для контроля этапов производства холерной химической вакцины и рассмотренными в данной работе. Отмечено, что применение клеточной линии СНО-К1 наиболее перспективно для замены биомоделей на промежуточных этапах контроля активных компонентов холерной химической вакцины. Разработанный методический подход впервые предлагается использовать на этапах производства холерной бивалентной химической вакцины. культура клеток, Vibrio cholerae, холерная химическая вакцина, контроль производства, холера. Vol 36(2020) N 3 p. 82-89; DOI 10.21519/0234-2758-2020-36-3-82-89A.V. Gaeva1*, O.V. Gromova1, O.S. Durakova1, S.V. Generalov1, L.F. Livanova1, O.A. Volokh1 Determination of Specific Activity of Cholera Chemical Vaccine Components using Cell Culture 1Russian Research Anti-Plague Institute «Microbe» of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Saratov, 410005 *[email protected] Received - 26.11.2019; Accepted - 15.05.2020 References The methods has been described to determine the dynamics of toxin production by the Vibrio cholerae 569B strain during submerged cultivation in bioreactor and of the antigenic activity of specific choleragen anatoxin fraction by anatoxin binding levels using cell cultures. High degree of consistency was observed between the results obtained via the method under consideration and those obtained via control methods at different stages of cholera chemical vaccine production. It was shown that the CHO-K1 cell line is the most promising substitute for biomodels at the intermediate stages of control of active cholera chemical vaccine components. The developed methodological approach was first proposed for use at the stages of cholera chemical bivalent vaccine manufacturing. cell culture, Vibrio cholerae, cholera chemical vaccine, production control, cholera.

The Natural Radioactivity in Food: A Comparison Between Different Feeding Regimes

2019 ◽  
Vol 15 (5) ◽  
pp. 493-499 ◽  
Author(s):  
Francesco Caridi ◽  
Santina Marguccio ◽  
Alberto Belvedere ◽  
Maurizio D`Agostino ◽  
Giovanna Belmusto
Keyword(s):  
Vegetable Oils ◽  
Natural Radioactivity ◽  
Specific Activity ◽  
Mean Value ◽  
Vegetable Food ◽  
Fish Samples ◽  
Feeding Regimes ◽  
Italian Origin ◽  
Dose Levels

Background: In this article a comprehensive study was carried out for the determination of natural radioactivity in animal and vegetable food (meat, fish, milk and derivates, legumes, cereals and derivates, fruit, hortalizas, vegetables, vegetable oils) typical of different feeding regimes, for the age category higher than 17 years. Methods: A total of eighty-five samples of Italian origin, coming from large retailers during the years 2014, 2015 and 2016, were analyzed through HPGe gamma spectrometry. Results: The specific activity of 40K was investigated and its mean value was found to be: (106.3 ± 6.9) Bq/kg for bovine, swine and sheep meat; (116.5 ± 9.7) Bq/kg for fish; (52.9 ± 3.1) Bq/kg for milk and derivates; (271.9 ± 16.7) Bq/kg for legumes; (67.2 ± 4.7) Bq/kg for cereals and derivates; (52.7 ± 4.4) Bq/kg for fruit; (72.9 ± 5.6) Bq/kg for hortalizas; (83.9 ± 6.5) Bq/kg for vegetables; lower than the minimum detectable activity for vegetable oils. For animal food the highest mean 40K activity concentration was found in fish samples; for vegetable food the highest one was detected in legumes. Conclusion: The evaluation of dose levels due to the food ingestion typical of Mediterranean, Vegetarian and Vegan diets was performed. The annual effective dose was found to be 0.16 mSv/y, 0.41 mSv/y and 0.54 mSv/y, respectively.

Determining the Norton’s Equivalent Model of Distribution System with Distributed Generation (DG) for Stability Analysis

2019 ◽  
Vol 12 (2) ◽  
pp. 190-198
Author(s):  
Sunny Katyara ◽  
Lukasz Staszewski ◽  
Faheem Akhtar Chachar
Keyword(s):  
Distributed Generation ◽  
Normal Condition ◽  
Distribution System ◽  
Distribution Networks ◽  
Equivalent Model ◽  
Stability Factor ◽  
Matlab Simulation ◽  
The Stability ◽  
Stability Issues

Background: Since the distribution networks are passive until Distributed Generation (DG) is not being installed into them, the stability issues occur in the distribution system after the integration of DG. Methods: In order to assure the simplicity during the calculations, many approximations have been proposed for finding the system’s parameters i.e. Voltage, active and reactive powers and load angle, more efficiently and accurately. This research presents an algorithm for finding the Norton’s equivalent model of distribution system with DG, considering from receiving end. Norton’s model of distribution system can be determined either from its complete configuration or through an algorithm using system’s voltage and current profiles. The algorithm involves the determination of derivative of apparent power against the current (dS/dIL) of the system. Results: This work also verifies the accuracy of proposed algorithm according to the relative variations in the phase angle of system’s impedance. This research also considers the varying states of distribution system due to switching in and out of DG and therefore Norton’s model needs to be updated accordingly. Conclusion: The efficacy of the proposed algorithm is verified through MATLAB simulation results under two scenarios, (i) normal condition and (ii) faulty condition. During normal condition, the stability factor near to 1 and change in dS/dIL was near to 0 while during fault condition, the stability factor was higher than 1 and the value of dS/dIL was away from 0.

Synthesis and solvatochromic studies of 2- amino-3-phenolazo1-(4-sulfophenyl)-3-methyi-5-pyrozolone and use it for the determination of trace amount of Nickel (II) in blood samples

2016 ◽  
Vol 5 (10) ◽  
pp. 4920
Author(s):  
Amar M. Ali ◽  
Hussain. J. Mohammed*
Keyword(s):  
Stability Constant ◽  
Trace Amount ◽  
Benzenesulfonic Acid ◽  
Determination Method ◽  
Blood Samples ◽  
Determination Of Nickel ◽  
Normal Human ◽  
The Stability ◽  
Normal Human Blood

A new, simple, sensitive and rapid spectrophotometric method is proposed for the determination of trace amount of Nickel (II). The method is based on the formation of a 1:2 complex with 4-(4-((2-hydroxy-6-nitrophenyl) diazenyl) -3-methyl-5-oxo-2, 5-dihydro-1H-pyrazol-1-yl) benzenesulfonic acid (2-ANASP) as a new reagent is developed. The complex has a maximum absorption at 516 nm and εmax of 1. 84 X 105 L. mol-1. cm-1. A linear correlation (0. 25 – 4. 0μg. ml-1) was found between absorbance at λmax and concentration. The accuracy and reproducibility of the determination method for various known amounts of Nickel (II) were tested. The results obtained are both precise (RSD was 1. 2 %) and accurate (relative error was 0. 787 %). The effect of diverse ions on the determination of Nickel (II) to investigate the selectivity of the method were also studied. The stability constant of the product was 0. 399 X 106 L. mol-1. The proposed method was successfully applied to the analysis of diabetes blood and normal human blood. 

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