Sorption capacity of polystyrene plates used in immunoenzyme analysis

The data indicating that the degree of sorption of the antigen onto the plate depends on the duration of this process are presented. As shown in this investigation, the lower the adsorption activity of the plates, the longer the process of the sensitization of plates with the antigen. The process of the sorption of the antigen onto the plates with low adsorption capacity may be accelerated by sensitizing the plates at different temperatures. Besides, the adsorption capacity of the plates was increased by ultraviolet irradiation.     

Method of determining proteolytic activity by using a conjugate of bovine serum albumin with peroxidase

The authors propose a method for determination of proteolytic activity, based on the hydrolysis of peroxidase-labeled molecules of bovine serum albumin immobilized on the surface of polystyrene microassay plates with the subsequent determination of peroxidase activity on the carrier or in the solution. The optimum conditions for the sorption of the labeled substrate have been established. The method permits the determination of bacillary alkaline protease at a concentration of 01. microgram/ml within 45 minutes. The determination of four proteases has demonstrated that this method shows good correlation with the routine one (r = 0.98), but is more sensitive and less time- and labor-consuming.     

The use of the method of the complete advisability function for evaluating the quality and standardization of the plates in solid-phase EIA

An attempt to use the method of the complete advisability function for the evaluation of the quality of plates and their standardization in the enzyme-linked immunosorbent assay (ELISA) has been made. Correlations have been empirically established, thus making it possible to calculate the optimum proportions of different ELISA factors: the dose of the antigen (antibody) and the time of sorption, which ensures the best ELISA results with a given type of plates.     

Immunoenzyme analysis using paper disks

Filter paper discs have been used in the enzyme immunoassay (EIA) as solid phase instead of polystyrene plates. The use of paper discs has made it possible to achieve a multiple increase in the sensitivity of sandwich EIA, thus permitting the detection of Yersinia pestis capsular antigen at a concentration of 0.4 ng/ml. Paper discs can be used not only for the sorption of antigen and antibodies, but also for the affinity purification of preparations.     

The sorption of a protein-polysaccharide complex isolated from Neisseria meningitidis serogroup B on aluminum hydroxide gels and the immunological activity of the sorbed preparations

The protein-polysaccharide complex, isolated from group B N. meningitidis, is a variant of vaccine for the prophylaxis of group B N. meningitidis infection. In this investigation the influence of the complex of the physical properties of aluminium hydroxide gels, the amount of gel, pH and the duration of sorption on the process of sorption has been studied. Aluminium hydroxide has been shown to produce a stimulating effect on the response of mice to the polysaccharide and protein contained in the complex after immunization made in two injections. Gels with a smaller particle size have been found to possess greater adjuvant activity, as well as greater absorbing activity. The immunological activity of the complex, adsorbed ex tempore, has proved to be no different from that of the complex adsorbed in an hour.     

Comparative characterization of the polystyrene plates used in enzyme immunoassay

The data on the comparison of the quality of polystyrene plates manufactured in the USSR with those manufactured by a number of foreign producers are presented. These data indicate that the capacity of antibodies adsorbed on a polystyrene plate for the sorption of antigens depend on the properties of the polymer surface and vary for different plates.     

Study of water sorption on modified Agave fibres

Polymer composite materials are usually reinforced by synthetic matter such as carbon or glass fibres. However, owing to their good mechanical properties and low density, natural fibres are now increasingly being considered as reinforcement. With the aim of a new natural fibre based composite, various chemical treatments have been performed on Agave (Americana L.) fibres in order to improve their compatibility with the polymer matrix and to reduce their affinity for water. The effect of these treatments on the fibre water sorption power has been investigated by means of a micro-balance. Equilibrium water sorption isotherms have been deduced from weight variations of the fibres under water vapor pressure increments. Several specific physico-chemical models have been tested to describe the water sorption isotherms. The Park’s model was found to describe the experimental results accurately and over a wide activity range. The sorption kinetics was also exploited in order to evaluate the diffusivity of water in the fibres. The variation of the water diffusion coefficient with water concentration is in agreement with the triple sorption mode described by the Park’s model. These results show a global increase of moisture resistance of the fibres after chemical treatment. This effect is interpreted in terms of chemical and structural modifications of the cell-wall structure.     

Study of the sorption of glycopeptide antibiotics on an ultradisperse carbon sorbent

The sorption capacity of a novel ultradisperse carbon sorbent (UDCS) towards antibiotics of the glycopeptide group, namely, eremomycin, vancomycin, ristomycin A, and teicoplanin A2, has been studied. The conditions for maximum sorption of the antibiotics from solutions have been determined, and a mathematical model of the sorption of the antibiotics of the above named group has been put forward.     

Interrelationships between Water and cellular metabolism in Artemia cysts. VIII Sorption isotherms and derived thermodynamic quantities

An analysis of water vapor sorption by cysts of the brine shrimp, Artemia salina, has shown that at environmental water activities (aw) of 0.95 or less, the cysts equilibrate with the aw of their environment. Above this aw the metabolic activity of the cysts participates directly in their water content, and equilibration does not occur. In contrast, dried cysts killed by heat treatment or exposure to ammonia fumes equilibrated with all values of aw examined. Analysis of the temperature dependence of sorption isotherms revealed that below cyst hydrations of about 0.3 g H2O/g dried weight the temperature coefficient for water sorption was negative, but became positive at hydrations appreciably in excess of this value. Estimates for the differential and integral net enthalpy and entropy changes accompanying the sorption of water have been calculated from isotherms. These results have been interpreted and integrated with those from previous work on the hydration-dependence of metabolic activity. All of the examined hydration properties of the cysts have been shown to be due chiefly to the cellular component, and not the acellular shell. Analysis of the data by the Bradley equation has shown that the hydration behavior of the shell obeys this relationship, whereas that of the cellular component does not.     

Structural Relaxation During Drying and Rehydration of Food Materials��the Water Effect and the Origin of Hysteresis

The state of water in foodstuffs is a guiding principle in food design, and the equilibrium concept of water activity (Aw) is ubiquitous. It is regarded as a primary variable or “hurdle” in preservation technology, and a key variable influencing chemical reaction during storage. However, the amount of water in any system differs as function of water activity depending whether it is determined by water sorption or desorption. Even though this hysteresis behaviour has already been described in the literature, no physical interpretation of its origin has yet been proposed with respect to detailed molecular organisation. This work shows, for two different food powders, gluten and a milk-based product that the hysteresis disappears when either go through their glass transition. A more complete DSC analysis for gluten during different sorption/desorption cycles demonstrates that the hysteresis is dependent on the ageing of the material, which evolves in the glassy state and is induced by structural relaxation.     

Sorption von Cellulase an Weizenstroh und seine Komponenten

By comparing different activity data of the buffered cellulase solution before and after contact with the substrate the interaction between Penicillium janthinellum cellulase and wheat straw, resp. its components (holocellulose and isolated lignin) has been investigated. The loss of activity due to sorption or denaturation has been found to differ widely between the different activity data and between the various substrates. A remarkable loss of enzyme activity was observed after contact with isolated straw lignin. The differences in activity decrease between the cellulose and the lignin moiety were found to be largent with the cellobiase activity.     

The hydration of proteins in nearly anhydrous organic solvent suspensions

Water sorption isotherms at 27°C have been measured for lysozyme and chymotrypsin in suspensions of toluene, di(n-butyl) ether, n-propanol, and a solution of 1M n-propanol in benzene. Sorption isotherms for the different suspensions are compared by converting solvent water content to the thermodynamic activity of water in each solvent. The sorption behavior is also compared to that for the two proteins hydrated from the vapor phase. At low water activities, all sorption isotherms are similar when compared on the basis of water activity. However, at higher activities, water sorption by the proteins in the organic suspensions is suppressed relative to the sorption of water vapor. The greatest suppression is observed for n -propanol, which suggests that the suppression may be due to a competition for water-binding sites on the protein by the organic solvent. Sorption isotherms at low water activities have also been predicted using a thermodynamic model in which it is assumed that water binds selectively to the ionizable residues on the surface of the protein. A comparison of predicted and measured sorption isotherms shows that the model can provide reasonable estimates of water sorption in nonpolar or moderately polar organic solvent suspensions at low levels of hydration. © 1993 John Wiley & Sons, Inc.     

Heavy metals biosorption from aqueous solution by simple eucaryotic organisms

Biosorption of cadmium and chromium (III) ions by means of selected yeast species has been estimated. Kinetics and equilibrium measurements have shown the reliable efficiency of both metals removal for Candida tropicalis. The influence of pH and ionic strength on biosorption process has been examined as well. For both metals the adsorption isotherms have been presented. The equilibrium of chromium (III) sorption has appeared compatible to Langmiur model and the maximum sorption capacity has been determined.     

Identification of a Cholinergic-Specific Antigen Chol-1 as a Ganglioside

Abstract: An antiserum specific for cholinergic terminals was used to identify an antigen conserved between Elasmobranchs and mammals. Immunohistochemistry and a cytotoxicity test were used to assay the binding of antibody to mammalian terminals. Torpedo electric organ gangliosides totally abolished antibody binding. The highest inhibitory activity was associated with a single polysialoganglioside band on TLC plates. Neuraminidase altered the migration of the inhibitory activity on TLC plates. Antibody binding was inhibited by ganglioside fractions derived from chicken and mammalian brains. A summary of those tissues in which the antigen has been detected is presented. The possible function of the antigen is discussed.     

Pseudo-isotherms for the sorption of cadmium ion onto tree fern

The kinetics of cadmium sorption on to tree fern has been investigated based on the assumption of a pseudo-second order rate law. The batch sorption model has been applied to predict the rate constant of sorption and the equilibrium capacity with the effect of initial cadmium concentration. The experimental results have been analysed by pseudo-Langmuir, pseudo-Freundlich and pseudo-Redlich–Peterson isotherms based on pseudo-second order kinetic expression constant. Both pseudo-Langmuir and pseudo-Redlich–Peterson isotherms were found to represent the measured sorption data well. According to the evaluation using the pseudo-Langmuir equation, the monolayer sorption capacity was obtained to he 16.3 mg/g. In addition, χ² analysis was also used to determine the most suitable model. Pseudo-Langmuir and pseudo-Redlich–Peterson were also the best models for the experiment data from χ² analysis.     

Drug release and its relationship with kinetic and thermodynamic parameters of drug sorption onto starch acetate fibers

Drug release and its relationship with kinetic and thermodynamic parameters of drug sorption onto starch acetate (SA) fibers have been studied using Diclofenac, 5‐Fluorouracil (5‐Fu), and Metformin as model drugs. The sorption method is more flexible and can avoid limitations or problems which occur with molten or dissolution methods. To understand drug release of sorption loading, kinetic and apparent thermodynamic parameters, such as diffusion coefficient, activation energy for diffusion, affinity, and sorption enthalpy and entropy, have been investigated. The quantitative relationship between drug release and drug‐loading concentration, affinity, and activation energy for diffusion has been established to predict the initial burst and subsequent release of the drugs. Up to 12% of Diclofenac, based on the weight of SA, can be loaded onto fibers using the sorption method. Drugs with higher activation energy for diffusion, lower diffusion coefficients, and higher affinity for SA fiber, such as Diclofenac, are more suitable for sorption loading. It has also been found that elevated temperatures will achieve higher loading capacity and a more constant release rate. Biotechnol. Bioeng. 2010. 105: 814–822. © 2009 Wiley Periodicals, Inc.     

Adaptation of a gamma-glutamyl transpeptidase assay to microtiter plates

A kinetic assay for measuring gamma-glutamyl transpeptidase (GGT) activity has been adapted to microtiter plates and an automated microtiter plate reader. This method permits the simultaneous analysis of enzyme activity in a large number of samples incubated with the chromogenic GGT substrate gamma-glutamyl-p-nitroanilide. A major advantage of this assay over previously reported methods is the substantial reduction in the time needed for measuring sample enzyme activity. In addition, reduction of the total assay volume to 0.28 ml conserves both sample and reagents. This method has been calibrated at 23 degrees C using purified GGT, and used to analyze GGT activity in human sera. The assay is sensitive over a range of 3-200 U/liter.     

A rapid TLC bioautographic method for the detection of acetylcholinesterase and butyrylcholinesterase inhibitors in plants

A simple and rapid bioautographic enzyme assay on TLC plates has been developed for the screening of acetylcholinesterase and butyrylcholinesterase inhibition by plant extracts. Enzyme activity was detected by the conversion of naphthyl acetate into naphthol and the formation of the corresponding purple-coloured diazonium dye with Fast Blue B salt. Inhibitors of cholinesterases produced white spots on the dye-coloured background of the TLC plates. The alkaloids galanthamine and physostigmine, which are known inhibitors of acetylcholinesterase, were used to determine the sensitivity of the assay. Various plant extracts were tested using the bioassay.     

Macrokinetics of enzyme sorption on porous beads accompanied by complex formation with an immobilized ligand

A mathematical model has been proposed for enzyme sorption on porous beads accompanied by formation of a stable complex with an immobilized ligand. It has been experimentally verified by using the system trypsin (EC 3.2.21.4) - immobilized bovine basic polyvalent trypsin inhibitor on porous silica gel. The experimental results for kinetics of the non-specific/specific trypsin sorption on a carrier agree with the model. The value of the coefficient of trypsin diffusion in macroporous silica gel was calculated.     

A telomerase enzymatic assay that does not use polymerase chain reaction, radioactivity, or electrophoresis

A telomerase assay has been developed for high-throughput screening in 96-well microtiter plates. A crude cell lysate which adds telomere repeats to a biotinylated DNA primer is the source of telomerase. The telomerase-extended primer is hybridized to a digoxigenin-labeled telomere antisense DNA probe. The hybrid is further processed by enzyme-linked immunosorbent assay (ELISA) as follows. The biotinylated hybrid is captured on streptavidin-coated microtiter plates. The immobilized hybrid is probed with alkaline phosphatase-antidigoxigenin and detected via chemiluminescent readout. The limit of detection of a chemically synthesized tetra-telomere repeat was about 10 attomoles. Apparent telomerase activity was detected in lysates of 293T cells. The signal to background for the assay (ratio of signal for the complete assay mixture divided by the signal for the assay mixture without primer) was around 10. An automated system that performed unattended runs of up to 17 96-well microtiter plates in 8h was constructed.