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Investigation of plasma protein adsorption on functionalized nanoparticles for application in apheresis.

Particles with specific ligands for the adsorption of plasma proteins can be used in therapeutic or preparative apheresis. The development of these particles may benefit from an improved knowledge of the relationship between protein adsorption and the structure of ligands. Nanoparticles were functionalized with aliphatic diamines of increasing chain length; with the amino acids lysine, tryptophan, histidine, and their corresponding amines; and with tryptophan and histidine spaced with diamines of different length. Suitable protocols were developed for the washing of particles and the subsequent desorption of proteins adsorbed from human plasma. The adsorption pattern, as well as the quantification of the overall adsorption of proteins on these modified particles, was investigated with gel electrophoresis. This was followed by immunoblotting which yielded specific assessments of bound human serum albumin and fibrinogen. The comparison of protein adsorption with surface charge density and measured hydrophobicities yielded no simple correlations although in general more hydrophobic ligands bound higher quantities of protein. The detection of human serum albumin yielded similar results because it was observed for overall protein adsorption while the adsorption of fibrinogen expressed a different pattern. In this case, particular nanoparticles functionalized with aliphatic diamines bound significantly higher amounts of fibrinogen than all other ligands.

Adsorption↗

Anti-Vel reactivity diminished by adsorption with rabbit RBC stroma.

BACKGROUND: An anti-Vel, nearly missed in antibody identification studies, and the effect of a commercially available rabbit RBC stroma (RESt, Immucor) adsorptions on eight anti-Vel sera are reported. Anti-Vel is an antibody to an antigen of high prevalence. CASE REPORT: A 48-year-old woman with chronic vaginal bleeding presented with a Hct of 14.7 percent. The transfusion service was not informed of her history of anti-Vel when she was transferred from another institution. Studies performed on an emergency request for transfusion were interpreted as a cold autoantibody as adsorption with a commercial source of RESt eliminated the reactivity. Stored anti-Vel sera were tested by titration studies before and after adsorption with commercial RESt. RESULTS: Serum from the index case did not react after adsorption with RESt at the transfusion service. Studies with the stored anti-Vel indicated antibody adsorption with four of four samples at immediate spin (IS) and room temperature (RT) phases, four of eight samples at 37 degrees C in albumin (ALB) phase, and four of eight samples at ALB-IgG-AGT phase. Variations in antibody reactivity were observed in the samples tested, but RESt adsorption diminished antibody reactivity in most samples. All eight stored sera demonstrated some reactivity in at least one phase after adsorption with RESt. CONCLUSION: Anti-Vel was completely or partially adsorbed by RESt. Caution should be used when interpreting cold agglutinins with this method. The manufacturer warns that uncommon alloantibodies may be adsorbed.

ABO Blood-Group System↗

The effect of food and ice cream on the adsorption capacity of paracetamol to high surface activated charcoal: in vitro studies.

The effect of added food mixture (as if food was present in the stomach of an intoxicated patient) or 4 different types of ice cream (added as a flavouring and lubricating agent) on the adsorption of paracetamol (acetaminophen) to 2 formulations of activated charcoal was determined in vitro and compared with results from previous investigations showing a maximum adsorption capacity to the two activated charcoal-water slurries at about 0.62-0.72 g paracetamol/g activated charcoal. Activated charcoal (Carbomix or Norit Ready-To-Use), simulated gastric (pH 1.2) or intestinal (pH 7.2) fluid, and paracetamol were mixed with either food mixture or ice cream followed by one hr incubation. The maximum adsorption capacity of paracetamol to activated charcoal was calculated using Langmuirs adsorption isotherm. Paracetamol concentration was analyzed using high pressure liquid chromatography. In the presence of food, the paracetamol adsorption capacity of the 2 activated charcoals was reduced by max. 19% (P<0.05) for Carbomix(R) and by max. 11% (P<0.05) for Norit Ready-to-use compared to control without food (Hoegberg et al. 2002). Depending on which type of ice cream was mixed with the charcoal, the reductions compared to control (Hoegberg et al. 2002) varied between 11% and 26%. Even though a reduction in drug adsorption to activated charcoal was observed when food mixture or ice cream was added, the remaining adsorption capacity of both types of activated charcoal theoretically was still able to provide an effective gastrointestinal decontamination.

Acetaminophen↗

Conformation and orientation of a protein folding intermediate trapped by adsorption.

Although adsorption-induced conformational changes of proteins play an essential role during protein adsorption on interfaces, detailed information about these changes is lacking. To further the current understanding of protein adsorption, in this study, the orientation, conformation, and local stability of bovine alpha-lactalbumin (BLA) adsorbed on polystyrene nanospheres is characterized at the residue level by hydrogen/deuterium exchange and 2D NMR spectroscopy. Most of the adsorbed BLA molecules have conformational properties similar to BLA molecules in the acid-induced molten globule state (A state). A folding intermediate of BLA is thus induced and trapped by adsorption of the protein on the hydrophobic interface. Several residues, clustered on one side of the adsorbed folding intermediate of BLA, have altered amide proton exchange protection factors compared to those of the A state of BLA. This side preferentially interacts with the interface and includes residues in helix C, the calcium binding site, and part of the beta-domain. Local unfolding of this interacting part of the adsorbed protein seems to initiate the adsorption-induced unfolding of BLA. Adsorption-induced protein unfolding apparently resembles more the mechanical unfolding of a protein than the global unfolding of a protein as induced by denaturant, pH, or pressure. 2D macromolecular crowding prevented the minority of adsorbed BLA molecules, which arrived late at the interface, to unfold to the A state. Protein adsorption is a novel and challenging approach to probe features of the free energy landscapes accessible to unfolding proteins.

Adsorption↗

Kinetic and structural characterization of adsorption-induced unfolding of bovine alpha -lactalbumin.

Conformational changes of bovine alpha-lactalbumin induced by adsorption on a hydrophobic interface are studied by fluorescence and circular dichroism spectroscopy. Adsorption of bovine alpha-lactalbumin on hydrophobic polystyrene nanospheres induces a non-native state of the protein, which is characterized by preserved secondary structure, lost tertiary structure, and release of calcium. This partially denatured state therefore resembles a molten globule state, which is an intermediate in the folding of bovine alpha-lactalbumin. Stopped-flow fluorescence spectroscopy reveals two kinetic phases during adsorption with rate constants k(1) approximately 50 s(-1) and k(2) approximately 8 s(-1). The rate of partial unfolding is remarkably fast and even faster than unfolding induced by the addition of 5.4 m guanidinium hydrochloride to native alpha-lactalbumin. The large unfolding rates exclude the possibility that unfolding of bovine alpha-lactalbumin to the intermediate state occurs before adsorption takes place. Stopped-flow fluorescence anisotropy experiments show that adsorption of bovine alpha-lactalbumin on polystyrene nanospheres occurs within the dead time (15 ms) of the experiment. This shows that the kinetic processes as determined by stopped-flow fluorescence spectroscopy are not affected by diffusion or association processes but are solely caused by unfolding of bovine alpha-lactalbumin induced by adsorption on the polystyrene surface. A scheme is presented that incorporates the results obtained and describes the adsorption of bovine alpha-lactalbumin.

Adsorption↗

Surface exclusion and molecular mobility may explain Vroman effects in protein adsorption.

Data on protein adsorption usually show that for increasing surface coverage the adsorption velocity decreases much faster than linearly. This contrasts to the classical Langmuir model with an adsorption velocity proportional to the number of unoccupied binding sites. It has been shown that this non-linearity may explain phenomena like transient adsorption of different proteins from a protein mixture or dilution-dependent changes in binding properties, collectively called Vroman effects. However, the molecular mechanisms explaining this non-linear behavior remain to be established. A Monte Carlo simulation model is presented that incorporates steric hindrance, lateral mobility and mutual interactions of adsorbed molecules. Experimental data on the adsorption kinetics of prothrombin and annexin V, a recently discovered anticoagulant protein, at phospholipid bilayers are analyzed with this model. A major conclusion is that the steep decline in adsorption rates for increasing surface coverage can be explained, without assuming repulsive forces between adsorbed molecules, as a surface exclusion effect combined with lateral mobility of adsorbed molecules. The fact that annexin V shows this effect to a much lesser degree than prothrombin is tentatively explained by clustering of adsorbed annexin V molecules. A qualitative effect of lateral mobility on the adsorption characteristics, predicted by the model, is confirmed in experiments in which the fluidity of the bilayers was manipulated.

Adsorption↗

An investigation of the adsorption of organic dyes onto organo-montmorillonite.

Adsorption of organic dyes, crystal violet (CV), orange II (OR), and phenol red (PR), onto organo-clay was investigated in a batch type reactor at 25 degrees C. The organo-clay was obtained by modifying montmorillonite with a cationic surfactant, cetylpyridinium (CP), and used as an adsorbent. We conducted experiments to find out the effect of pH and solvent on the adsorption affinity of organic dyes for the modified montmorillonite. From the results, we observed that the adsorption capacity on the organo-montmorillonite decreased in the order CV > OR > PR at all pH values examined (pH 3, pH 7, and pH 11). It mostly resulted from the difference in solubility and the molecular weight of the solutes. In a 30-V/V % methanol/water cosolvent solution, the adsorption capacity of the dyes decreased compared to that in aqueous solution. In addition, the adsorption capacities of OR and PR on CV-montmorillonite were lower than those on CP-montmorillonite. These results might show that partitioning by CP was superior to the adsorption by CV to hold the solute molecules on the surface of montmorillonite. The Langmuir and Redlich-Peterson (RP) models were used to represent the adsorption equilibria of the organic dyes.

Adsorption↗

Adsorption of an amine drug onto microcrystalline cellulose and silicified microcrystalline cellulose samples.

The adsorption of a model amine drug (tacrine hydrochloride) from aqueous solution onto 21 microcrystalline cellulose (MCC) based samples has been investigated. The MCC source (manufacturer) affected adsorption. The adsorption appeared to be fully reversible. Adsorption was reduced by the use of high-density grade MCC, high-energy milling, and silicification. Adsorption of the model drug was not affected by the particle size of the MCC. Significant variations of the adsorption characteristics between batches of certain MCC products were found. The primary mode of adsorption was by ion exchange.

Adsorption↗

Kinetics of adsorption and uptake of Cu2+ by Chlorella vulgaris: influence of pH, temperature, culture age, and cations.

Adsorption and uptake of Cu2+ by Chlorella vulgaris were distinguished by extracting the surface-bound Cu2+ with EDTA. The uptake of Cu2+ followed Michaelis Menten kinetics. The maximum rate of Cu2+ uptake (0.362fmolcell(-1) h(-1)) was obtained at pH 6.0. The rate of Cu2+ uptake was greater for cultures in the exponential phase of growth, and increased with a rise in temperature from 6 to 25 degrees C, thus pointing towards an active mechanism. The maximum number of Cu2+ binding sites was 3.245 fmol cell(-1) at pH 4.5. Adsorption of Cu2+ was strongly pH-dependent thereby indicating that the number and nature of metal binding sites on the cell surface change with changing chemistry of the solution. Unlike uptake, the adsorption remained unaffected by small changes in temperature. Older cultures displayed a higher Cu2+ adsorption capacity than the exponentially growing ones thus suggesting generation of new and/or additional Cu2+ binding sites on older cells of C. vulgaris. By pH titration, the cation-exchange capacity of Chlorella, measured in terms of H+/ Na+ exchange, was about 17 fmol cell(-1) at pH 10.5. Negligible cation exchange capacity at and below pH 5.0 indicated that ion exchange was not the sole mechanism of Cu2+ adsorption by Chlorella. The uptake and adsorption of Cu2+ were inhibited by 100 microM of various cations including other heavy metal ions. The general concept that cations competitively inhibit accumulation of metals in living organisms does not hold for C. vulgaris. Non-competitive, uncompetitive and mixed inhibition of Cu2+ uptake and adsorption by various cations were more common than competitive inhibition.

Adsorption↗

Adsorption behavior of zinc and cadmium ion on granular activated carbon in singular and binary systems and the influence of nitrilotricetic acid as a complexing agent.

Adsorption characteristics of zinc and cadmium ion on granular activated carbon have been studied in dynamic and equilibrium aspects and singular and binary adsorption behavior were compared. Based on the van't Hoff equation, reaction orders and rate constants were observed to increase as initial concentration of adsorbate was raised for both metal ions. Adsorption of Zn and Cd ion was found to increase with temperature, and this endothermic nature of adsorption reaction was further verified by thermodynamic calculation. Taking nitrilotriacetic acid (NTA) as a ligand, the influence of complexing agent was examined and the variation of adsorption percentage was found to closely relate with the change in ionic form of NTA depending upon pH conditions and amount of complexing agent used. Features of binary adsorption are discussed for several influential parameters and experimental observations for both ions were correlated with a predicted adsorption isotherm based on a Langmuir multi-component model.

Adsorption↗

Contribution of cell outer membrane and inner membrane to Cu2+ adsorption by cell envelope of Pseudomonas putida 5-x.

The role of outer and inner membrane in the Cu2+ adsorption process by gram-negative bacterium Pseudomonas putida 5-x, which was isolated from local electroplating effluent with high Cu2+ accumulating capability, was studied. The results indicate that both the outer and inner membrane exhibited high Cu2+ adsorption capacity. Outer and inner membrane contributed about 30-35% and 20-25% parts of adsorption capacity in Cu2+ adsorption by cell envelope of Pseudomonas putida 5-x, respectively. The total contribution of outer and inner membrane to Cu2+ adsorption by cell envelope was much greater than that of peptidoglycan layer. The relatively high phospholipid content in the outer membrane might result in its greater heavy metal ions adsorption capacity. The Cu2+ binding process by the outer and inner membrane of Pseudomonas putida 5-x is the adsorption processes and can be described with Freundlich isotherms.

Adsorption↗

Adsorption of methabenzthiazuron on six allophanic and nonallophanic soils: effect of organic matter amendment.

This article reports on methabenzthiazuron [1-(1,3-benzothiazol-2-yl)-1,3-dimethylurea] (MBT) adsorption process on six agricultural allophanic and nonallophanic soils. The effect of amendment with exogenous organic matter was also studied. Adsorption kinetic fits an hyperbolic model. MBT adsorption reached an apparent equilibrium within 2 h and followed a second-order reaction. The maximum adsorbed amounts for natural soils ranged from 32 to 145 microg g(-1). Rate constants were considered relatively low (0.27-1.5 x 10(-4) [microg g(-1)](1-n) s-1); the slow process was attributed to a combined effect of difussion and adsorption. MBT adsorption fits the Freundlich model with r values > or =0.998 at P < or = 0.001 significance levels. Kf and Freundlich exponents (l/n) ranged from 5.3 to 82.1 cm3 g(-1) and from 0.66 to 0.73, respectively. Kf values for soils with a low organic matter content were lower than that obtained from the only typical allophanic soil derived from volcanic ash under study. Lineal regression analysis between Kf and organic matter content of nonallophanic soils gave a correlation coefficient of 0.980 (P = 0.02). Dispersion of Kd values together with close values of K(OM) indicate that organic matter (OM) was the principal component responsible for MBT adsorption in unamended soils. Addition of peat decreased soil pH and increased adsorption capacity for allophanic and nonallophanic soils. Kinetic experiments showed enhancements of Xmax values and lower rate constants.

Adsorption↗

The action of sphingomyelinase of Bacillus cereus on bovine erythrocyte membrane and liposomes. Specific adsorption onto these membranes.

Sphingomyelinase of Bacillus cereus was specifically adsorbed onto sphingomyelin liposome in the presence of Ca2+ or with the coexistence of Ca2+ and Mg2+, but not onto the liposome of phosphatidylcholine. In the presence of Ca2+, the enzyme adsorption onto bovine erythrocytes and liposome increased with an increase in the amount of sphingomyelin. These results support that the major binding site for sphingomyelinase in the erythrocytes is sphingomyelin. The temperature-dependence of enzyme adsorption was not influenced by a change in ATP content of bovine erythrocytes. After treatment of red cells with neuraminidase or pronase, enzyme adsorption at 0 degrees C lower than that at 37 degrees C was observed. In unsealed or right-side-out ghosts, the difference between the enzyme adsorption at 37 degrees C and that at 0 degrees C became less pronounced than in the erythrocytes. Furthermore, the extent of enzyme adsorption onto sphingomyelin liposome at 0 degrees C was almost equal to that at 37 degrees C. The enzyme adsorption onto the erythrocyte membrane and liposome was always enhanced in the presence of Ca2+; in the presence of Mg2+ alone, adsorption was observed only for erythrocyte ghosts and the adsorbed enzyme was released from the membrane after extensive degradation of sphingomyelin by sphingomyelinase. With the coexistence of Ca2+ and Mg2+, the enzymatic hydrolysis of sphingomyelin proceeded rapidly for the attack against liposome and all the membranes tested, whereas in the presence of Mg2+ alone, hydrolysis was observed only for the action of liposome and ghosts. No appreciable hydrolysis of sphingomyelin was observed in the presence of Ca2+ nor in the absence of divalent metal ions.

Adenosine Triphosphate↗

Removal of low-density lipoprotein from plasma by adsorption increases bradykinin and plasma nitric oxide levels in patients with peripheral atherosclerosis.

Low-density lipoprotein (LDL) adsorption using a dextran sulfate cellulose column is brought about by electrostatic binding between the positive charges of apolipoprotein B in LDL and the negative charges of dextran sulfate cellulose. There is general agreement that the initial contact phase in the coagulation pathway may be activated by a negatively charged surface such as dextran sulfate cellulose, resulting in the generation of bradykinin. We investigated whether the increase in the generation of bradykinin during LDL adsorption is accompanied by the activation of endogenous production of nitric oxide (NO) in patients with peripheral atherosclerosis. LDL adsorption therapy was repeated ten times over a period of 3 months in ten peripheral atherosclerosis patients. Treatment significantly reduced serum total cholesterol and LDL cholesterol levels. This was associated with a significant improvement in Fontaine's classification and ankle pressure index. We also measured the kinin-kallikrein system and plasma levels of NO in the same patients. The results showed that coagulation factors of the intrinsic pathway including high-molecular-weight kininogen and prekallikrein decreased markedly after initial adsorption compared with the levels before treatment. There was a marked increase in bradykinin and NO concentrations after the initial adsorption, compared with their levels before adsorption. Our results suggest that the generation of bradykinin and increased plasma levels of NO may contribute to the improvement in peripheral circulation after LDL adsorption in peripheral atherosclerosis patients.

Adsorption↗

The adsorption of argon, krypton, and xenon on activated charcoal.

Charcoal adsorption beds are commonly used to remove radioactive noble gases from contaminated gas streams. The design of such beds requires the adsorption coefficient for the noble gas. Here an extension of the Dubinin-Radushkevich theory of adsorption is developed to correlate the effects of temperature, pressure, concentration, and carrier gas on the adsorption coefficients of krypton, xenon, and argon on activated carbon. This model is validated with previously published adsorption measurements. It accurately predicts the equilibrium adsorption coefficient at any temperature and pressure if the potential energies of adsorption, the micropore volume, and the van der Waals constants of the gases are known.

Adsorption↗

Use of L-rhamnose to study irreversible adsorption of bacteriophage PL-1 to a strain of Lactobacillus casei.

L-rhamnose has been found to be useful to investigate the process of irreversible adsorption of PL-I phage to its host bacterium, Lactobacillus casei ATCC27092. L-rhamnose inhibited phage adsorption to cells without inactivating free phages. Adsorption inhibition was correlated with the concentrations of L-rhamnose. The inhibitory effect of L-rhamnose on phage adsorption was of a competitive nature against host cells. Among other saccharides tested, L-fucose, L-mannose and D-ribose showed a slight degree of adsorption-inhibiting activity. In early stages of phage adsorption in a tris-maleate buffer, where the binding of phages to cells was still reversible, addition of L-rhamnose resulted in the partial desorption of phages from the cells to which they had adsorbed. However, the number of infective phages desorbed by L-rhamnose treatment gradually fell off as incubation continued, showing that the phages became firmly bound to the cells. Therefore, it is possible to determine the number of phages irreversibly adsorbed to cells by using this desorption technique with L-rhamnose. The process of irreversible phage adsorption, that is, the formation of phage-cell complexes from which no more infective phages could be desorbed, was dependent on temperature and strongly inhibited at 0 degrees C.

Adsorption↗

Transport and kinetics in synthetic and immunospecific adsorption columns.

Immunological or physicochemical adsorption can be used to remove specific proteins from plasma. The most common adsorption systems are based on the principle of affinity or adsorption chromatography. An adsorption column can be considered to be analogous to a packed bed, and transport is modeled by accounting for solute convection, accumulation, uptake, and axial dispersion within the bed. Adsorption kinetics and transport within particles of the packed bed need to be described. The solution of model equations is solute specific and generally requires numerical methods. The application of column adsorption systems to on-line therapeutic adsorption is constrained by flow considerations, which influence processing time; adsorbent capacity, which dictates column volume and hence design; and adsorbent stability, which may impact on patient safety.

Adsorption↗

Protein adsorption by artificial membrane materials under filtration conditions.

Elevated plasma levels of numerous low molecular weight proteins (LMWP) in renal insufficiency are likely to contribute to the uremic syndrome. Dialysis-related amyloidosis, caused by the accumulation of beta 2-microglobulin (beta 2M), has highlighted the need for a renal replacement therapy that allows the elimination of LMWP in addition to small solutes. Synthetic membrane materials employed under hemofiltration conditions proved to be most effective in lowering elevated beta 2M plasma levels. In addition to convection, protein adsorption to artificial membrane materials is an important mechanism for beta 2M removal. Using an in vitro setup, 12 commercially available hemofilters representing 11 different membrane materials were perfused with human blood containing 125I-labeled plasma proteins. Under filtration conditions, total protein adsorption ranged from 338-2,098 mg/m2 of membrane surface, and the fraction of adsorbed LMWP varied between 14-70% of total protein adsorption and was negatively correlated to total protein adsorption. beta 2M adsorption showed up to an 8-fold difference between membranes, and was negatively correlated with total protein adsorption and positively correlated with the adsorption of LMWPs.

Adsorption↗