Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “IMMOBILIZATION”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 37 records · Page 2Linked to original sources
PubMed1992

Cell growth on immobilized cell-growth factor; 4: Interaction of fibroblast cells with insulin immobilized on poly(methyl methacrylate) membrane.

Insulin was immobilized on the surface-hydrolyzed poly(methyl methacrylate) membrane and the growth acceleration of mouse fibroblast cells, STO, by the immobilized insulin was investigated. It was found that insulin remains immobilized on the surface of nonbiodegradable membrane and interacts specifically with receptors existing on the biological membrane of fibroblast cells. The growth acceleration by immobilized insulin was enhanced by introduction of a spacer arm between insulin and the immobilization matrix. The amount of receptor proteins present on the biological membrane of fibroblast cells after culturing with insulin, immobilized on nonbiodegradable polymer membrane, was much higher than that after culturing with free insulin, implying the suppression of down-regulation in the case of immobilized insulin.

Animals↗
PubMed2003

Syntheses of immobilized G protein-coupled receptor chromatographic stationary phases: characterization of immobilized mu and kappa opioid receptors.

Opioid receptors are members of the superfamily of G protein-coupled receptors (GPCRs). Liquid chromatographic stationary phases containing either the human mu or kappa opioid receptor subtypes have been developed to study the binding between the opioid receptors and their ligands. The receptors were obtained from Chinese hamster ovary cells stably transfected with human mu or kappa receptor subtypes. The receptors were isolated through partial solubilization with sodium cholate detergent, and the partially purified receptor complex was immobilized in the phospholipid analogue monolayer of an immobilized artificial membrane liquid chromatographic stationary phase. The resulting phase was packed into a glass column (1.8 x 0.5 (i.d.) cm) and used in the on-line chromatographic determination of drug/ligand binding affinities to the immobilized opioid receptors. Preliminary on-line binding studies employing frontal chromatographic techniques were conducted with the known mu antagonist (naloxone) and a kappa agonist (U69593). The calculated dissociation constants (Kd) were 110 nM for naloxone and 84 nM for U69593. The results indicate that the immobilized receptors retained their ability to specifically bind ligands and were active for 1200 column volumes applied over two weeks. Zonal chromatographic experiments were also conducted, and competitive displacements of the marker ligands were observed. The results suggest that the immobilized opioid receptor stationary phases can be used to qualitatively assess the binding affinities of compounds to the immobilized receptors and represent the first example of the use of immobilized GPCRs in a chromatographic system.

Animals↗
PubMed1983

Comparative studies of the binding of some ligands to human serum albumin non-covalently attached to immobilized Cibacron Blue, or covalently immobilized on Sepharose, by column affinity chromatography.

A comparative study of the ligand-binding properties of human serum albumin was performed by the technique of affinity chromatography with the protein attached to immobilized Cibacron Blue F3GA (Blue Sepharose), or covalently immobilized on Sepharose. The binding strength of octanoate, decanoate and dodecanoate is much weaker when human serum albumin is attached to immobilized Cibacron Blue, indicating that the binding sites for fatty acids are involved in the attachment of human serum albumin to immobilized Cibacron Blue. The results revealed additional alterations of the ligand binding when human serum albumin was attached to immobilized Cibacron Blue, involving sites outside of the binding domains of fatty acids. Thus the stereoselective binding of L-tryptophan was abolished, and the resolution of the warfarin enantiomers was impaired. However, the binding strength of warfarin and salicylic acid was rather close to the values observed with human serum albumin covalently immobilized on Sepharose. It is suggested that the availability of the binding sites for L-tryptophan, warfarin and salicylic acid is partially blocked by the complex between albumin and the dye without direct participation in the complex-formation. An alternative interpretation involves an allosteric mechanism brought about by complex-formation between serum albumin and the immobilized Cibacron Blue.

Anthracenes↗
PubMed2005

Characteristics of vitamin C immobilized particles and sodium alginate beads containing immobilized particles.

This paper reports the properties of vitamin C encapsulated sodium alginate beads prepared by an alternative approach. The alternative encapsulation process mainly involves immobilization of vitamin C in hydrated zinc oxide layers and encapsulation of prepared immobilized particles in sodium alginate bead. The immobilization of vitamin C in hydrated zinc oxide layers was achieved by a coprecipitation process. Fourier transform infrared (FTIR) spectroscopy showed that the vitamin C was found to be stable after its immobilization. X-ray diffraction (XRD) studies revealed that anionic vitamin C molecules are adsorbed between positively charged zinc hydroxide layers with a 1:1 layer sequence, since well-defined change in basal spacing was observed. Well-defined change in surface morphology was observed by scanning electron microscopy (SEM) when vitamin C immobilized particles are encapsulated in sodium alginate bead. The biological activity of vitamin C was retained, even after its immobilization which was confirmed by 4-dihydroxy-L-phenylalanine (L-DOPA) oxidase inhibition and free radical scavenging activity studies. The release rate of vitamin C from immobilized particles and beads was sustained through an ion exchange process. A higher amount of stable vitamin C was recovered from the bead when compared to neat vitamin C itself.

Alginates↗
PubMed1989

Autoantibodies to lactate dehydrogenase in serum identified by use of immobilized protein G and immobilized jacalin, a jackfruit lectin.

In this method for identifying autoantibodies to lactate dehydrogenase (anti-LDs) in serum, we used immobilized Protein G to bind IgG-complexed LD and immobilized jacalin to bind IgA-complexed LD, leaving non-complexed LD in solution. The non-complexed LD and total LD were kinetically measured. We report results as LD bound to immobilized Protein G and LD bound to immobilized jacalin. Using sera demonstrating IgG and IgA anti-LDs by immunoelectrophoresis (IEP), respectively, we optimized the method for incubation time and concentration of binding agents. We demonstrated concomitant binding of LD and greater than or equal to 98% of IgG and of LD and greater than or equal to 92% of IgA. For LD bound to immunobilized Protein G the detection limit was 10 U/L, within- and between-run CVs ranged from 2.9% to 9.1%, and values for normal sera were less than or equal to 3% of total LD. Results for LD bound to immobilized jacalin were similar. We tested 10 sera displaying aberrant LD electrophoretograms: In seven, LD bound to immobilized Protein G was increased (range: 26-99% of total LD), indicating IgG-complexed LD. This was confirmed by IEP, demonstrating IgG1,2, or IgG3 anti-LDs in these sera. In the other three sera, LD bound to immobilized jacalin was increased (range: 38-72% of total LD), indicating IgA-complexed LD. This was confirmed by IEP, demonstrating IgA anti-LDs in these sera. Evidently this method is an alternative to IEP for identifying anti-LDs in serum.

Autoantibodies↗
PubMed1979

Immobilization of aminoacylase by adsorption to tannin immobilized on aminohexyl cellulose.

The immobilization of aminoacylase (N-acylamino acid amidohydrolase, EC 3.5.1.14) was investigated by using tannin immobilized on aminohexyl cellulose. The most active immobilized aminoacylase was obtained when aminoacylase was adsorbed to the immobilized tannin in a weak alkaline medium containing sodium chloride and n-butanol at 37 degrees C. The activity of the immobilized tannin-aminoacylase complex per unit volume was five times higher than that of the DEAE-Sephadex-aminoacylase complex used for industrial production of L-amino acids in our plants. The half-life of the immobilized tannin-aminoacylase complex was 20 days under continuous operation at a high concentration of substrate; on the contrary, that of the DEAE-Sephadex-aminoacylase complex was 0.5 days.

Adsorption↗
PubMed1994

Coenzyme production using immobilized enzymes. III. Immobilization of glucose-6-phosphate dehydrogenase from bakers' yeast.

Glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ 1-oxidoreductase, EC 1.1.1.49) from Bakers' yeast was immobilized with the highest activity on polyacrylamide beads possessing carboxylic functional groups activated by a water-soluble carbodiimide. The optimal pH values for the catalytic activity of the soluble and the immobilized glucose-6-phosphate dehydrogenase were practically identical, lying between pH 9.0 and 9.2. The optimal temperature for both the soluble and the immobilized enzyme was about 50 degrees C. The apparent Km values of the immobilized enzyme were slightly higher than those of the soluble enzyme. The immobilization improved the stability of the enzyme in the pH range 6.0-9.0 at 45 degrees C. The operational stability of the immobilized glucose-6-phosphate dehydrogenase proved favorable in a column experiment during 37 days of operation.

CME-Carbodiimide↗
PubMedDate not supplied

[Immobilization of Penicillium vitale glucose-oxidase on aminosilochrome and properties of immobilized enzyme].

Penicillium vitale glucose-oxidase modified by means of the carbohydrate component oxidation is added covalently to aminoorganosylochrome. The activity of the immobilized preparations is 20-38% depending on the protein-carrier ration in immobilization. Comparison of some properties of native and immobilized glucose-oxidase showed that the rH optimum of the immobilized glucose-oxidase is slightly widened towards the alkaline regions; the immobilized glucose-oxidase possesses a considerably higher pH-stability at pH alkaline values; the immobilized glucose-oxidase preparations are characterized by a significantly greater thermostability: their thermoinactivation constant at 65 degrees C is 8-10 times lower than that of the native enzyme.

Drug Stability↗
PubMed1982

[Use of immobilization for the study of glyceraldehyde 3-phosphate dehydrogenase. Immobilized dimers of the enzyme].

The immobilized dimers of glyceraldehyde 3-phosphate dehydrogenase have been obtained after dissociation of the tetrameric enzyme molecule linked by one of the subunits with Sepharose 4B. The catalytic parameters (V, Km and pH-dependence of activity) of the immobilized dimers and tetramers of the enzyme are identical. The immobilized dimers are capable of reassociating with the enzyme subunits from solution and possess a higher stability a compared to soluble dehydrogenase. The type of the dimer interaction with the reagents causing the "half-of-the-site reactivity" effect suggests that the immobilized dimers of the yeast and rat skeletal muscle enzymes possess the catalytic activity and, besides, are capable to express cooperative interactions between the active centers of the both subunits. The immobilized dimers of yeast dehydrogenase are able to form hybrids with soluble modified dimers of the yeast enzyme and with those from other sources. A procedure for the enzyme isolation from tissue extracts based on hybridization between the immobilized and soluble dimers of homologous dehydrogenases have been developed.

Animals↗
PubMed1993

Cell growth on immobilized cell growth factor. 9. Covalent immobilization of insulin, transferrin, and collagen to enhance growth of bovine endothelial cells.

Bovine endothelial cells were cultivated on the surface of polyurethane membranes or tubes immobilized with insulin, transferrin, and/or collagen, and growth was measured. For the purpose of immobilization, amino groups were introduced on the membranes or tubes by glow-discharge treatment in the presence of ammonia gas. The proteins were coupled to the amino groups on the surfaces by using dimethyl suberimidate (DMS) or water-soluble carbodiimide (WSC). Growth of bovine endothelial cells was higher on immobilized insulin or transferrin using DMS than that using WSC. The cell growth was higher on immobilized insulin or transferrin than on free insulin or transferrin, respectively. Though immobilized collagen did not affect the cell growth, coimmobilization with insulin or transferrin brought about greater acceleration of cell growth. In addition, the immobilized collagen was indispensible to maintain the endothelial cells on the material surface for a long time. The endothelial cells grown in the polyurethane tubes, on which insulin and collagen are coimmobilized, kept a normal cobblestone-like appearance and maintained the ability to secrete prostacyclin for 9 months.

Adsorption↗
PubMed1998

Indirect immobilization of recombinant proteins to a solid phase using the albumin binding domain of streptococcal protein G and immobilized albumin.

Immobilization of proteins to a solid phase leads to denaturation of the adsorbed molecules which may subsequently affect biological interactions. However, for many applications maintenance of the native structure is desired. Therefore, an indirect immobilization system was developed, based on binding of the albumin binding domain (ABP) of streptococcal protein G to rat serum albumin (RSA) precoated on a solid phase (RSA-microtiter plates). Escherichia coli vectors were adapted for production of recombinant protein fused to ABP and the 6 X His-tag. The expressed ABP tag was found to form homodimers. Plasmon resonance was used to study the interaction between an ABP fusion protein and immobilized RSA. Apparent on- and off-rates were calculated using a model for a bivalent analyte (k(a1) = 3.37 x 10(4) M(-1) s(-1), k(d1) = 1.23 x 10(-4) s(-1)). Thus, the stability of the ABP-RSA interaction can be explained by a slow off-rate. This was confirmed by chase experiments in an ELISA format. The ABP-RSA interaction remained stable after addition of different albumins. This immobilization system was used for the development of an ELISA to detect antibodies against Borna disease virus protein p40. The use of RSA-microtiter plates for indirect immobilization of ABP fusion protein was shown to be superior to direct adsorption on plastic. To obtain maximal antibody binding ten times less antigen was needed for indirect immobilization compared to direct adsorption. The binding capacity of the RSA-microtiter plates was determined to be about 0.8 pmol of monomeric ABP protein.

Adsorption↗
PubMed1985

Immobilization of immunoglobulins on silica surfaces. Kinetics of immobilization and influence of ionic strength.

The kinetics of, and the influence of ionic strength on, the immobilization of rabbit immunoglobulin G (IgG) on different types of well-characterized silica surfaces were investigated. Adsorptive immobilization was compared with covalent attachment via thiol-disulphide exchange reactions. The amount of immobilized IgG on five different types of silica surfaces as a function of IgG concentration, at two different ionic strengths, was determined. The IgG-solid-surface interaction involved different types of interaction forces, depending on the surface chemistry of the solid surface. The solid-surface chemistry is an important parameter determining the immobilized amount of IgG. When conditions for covalent attachment of IgG to the surfaces were fulfilled, the IgG showed high affinity and the immobilized amount of IgG showed a fast saturation. Changes in ionic strength showed no significant influence on the kinetics of immobilization on these surfaces. The amount of covalently attached IgG was partially ionic-strength-dependent, indicating that adsorptive interactions were involved. The results are of fundamental interest for the development of new immunosensors based on surface-concentration-measuring devices.

Adsorption↗
PubMedDate not supplied

Relation between number of immobile spermatozoa and pregnancies obtained during a twenty-year follow-up period immobile spermatozoa and fertility.

The clinical fertility of 1077 men examined with semen analysis including detection of number of immobile spermatozoa during the years 1950-52 was studied twenty years later with a questionnaire replied by 785 (72.9%). There was a significant relation (p less than 0.01) between decreasing number of immobile spermatozoa and increasing chance of getting living children, but no relation to abortions and pathological pregnancies. Furthermore, decreasing number of immobile spermatozoa was correlated to shorter time-interval between attempt to impregnate and first pregnancy obtained (p less than 0.01). Mean values for those obtaining living children was 34.7% immobile spermatozoa compared with 38.2% for those not doing so. Over 80% immobile spermatozoa, fertility was considerably reduced, and the borderline between normal and reduced male fertility should therefore be defined to 80% immobile spermatozoa.

Abortion, Spontaneous↗
PubMed2006

To immobilize or not immobilize: that is the question.

For those practicing selective immobilization, the strongest protection from liability is to avoid aggravating or causing injury by withholding c-spine immobilization from a patient who needs it. That means applying c-spine decision tools as objectively and carefully as possible. Prehospital providers in areas that use those tools should be trained to adequately assess the cervical spine and receive regular continuing education to maintain adequate performance. Applied correctly, selective spinal immobilization can be a great tool for decreasing patient discomfort, encouraging reluctant patients to seek medical care and reducing patient-provider conflicts. Any system that chooses to selectively apply cervical spine immobilization to trauma patients must regularly review cases in which spinal immobilization is withheld to determine effectiveness of, and ensure adherence to, decision criteria. Patients deserve the best care we can give them, with or without immobilization.

Decision Making↗
PubMed1977

Immobilization of urea cycle enzymes. II. Characterization of immobilized argininosuccinate synthetase.

Argininosuccinate synthetase (EC 6.3.4.5) was immobilized on CNBr-activated Sepharose 4B. Properties of the immobilized enzyme are described and compared with those of the native enzyme. The immobilized enzyme was much more stable than the native enzyme at 37 degrees C. It was further stabilized in the presence of the assay reagents. The optimum pH of the immobilized enzyme shifted towards alkalinity (approximately 0.5 unit). The apparent Michaelis constants measured for the immobilized enzyme were not greatly different from those measured for the native enzyme. Urea formation from citrulline was confirmed in a continuous column reactor by the coimmobilized argininosuccinate synthetase, argininosuccinate lyase (EC 4.3.2.1), and arginase (EC 3.5.3.1).

Animals↗
PubMed1992

Enzyme-based high-performance liquid chromatography supports as probes of enzyme activity and inhibition: the immobilization of trypsin and alpha-chymotrypsin on an immobilized artificial membrane high-performance liquid chromatography support.

Immobilized artificial membrane (IAM) HPLC supports have been used to immobilize the enzymes alpha-chymotrypsin and trypsin. The enzymes were trapped in hydrophobic cavities on the support and were not covalently attached to the IAM surface. The resulting IAM-enzyme supports retained the hydrolytic activity of the immobilized enzymes: the IAM-trypsin support catalyzed the hydrolysis of N alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), and the IAM-alpha-chymotrypsin support (IAM-ACHT) catalyzed the hydrolysis of a number of substrates, including tryptophan methyl ester. The activities of both supports were decreased by known enzyme inhibitors and the activity of the IAM-ACHT was affected by changes in pH and temperature. When a substrate was chromatographed on an IAM-ACHT HPLC, the hydrolytic activity of the immobilized enzyme could be determined from the resulting substrate/product ratios. These data were obtained either directly from the IAM-ACHT chromatogram or from the chromatogram produced by a coupled column system. The results of this study indicate that IAM-immobilized alpha-chymotrypsin and trypsin can be used as chromatographic probes for the qualitative determination of enzyme/substrate and enzyme/inhibitor interactions.

Benzoylarginine Nitroanilide↗
PubMed1979

Use of immobilized enzymes in automated clinical analysis: determination of uric acid and glucose using immobilized enzymes in column form.

We studied the use of immobilized enzymes, covalently bound to alkylaminosilane derivative of porous glass, to automated clinical analysis on uric acid and glucose in blood, serum and urine. A microcolumn with an immobilized enzyme was prepared and used in an AutoAnalyzer I continuous flow system. Uricase (EC 1.7.3.3) from Candida utilis and glucose oxidase (EC 1.1.3.4) from Aspergillus niger were immobilized for the determination of uric acid and glucose, respectively. Hydrogen peroxide produced by these oxidases was colorimetrically determined using horse-radish peroxidase (EC 1.11.1.7) and a hydrogen acceptor in solution. Sensitivity and wash charactertistics of a column with immobilized enzyme, 1.5 mm of inner diameter and up to 40 mm in length, were satisfactory at an assay speed of 50 samples per hour. The results correlated well with those obtained by other well established methods utilizing the AutoAnalyzer system. The immobilized enzymes were sufficiently stable for at least two months of 2000 tests when used repeatedly. Clinical trials proved that this method is capable of replacing the soluble enzyme method, giving reliable and reproducible results at lower cost.

Autoanalysis↗
PubMed2005

The covalent immobilization of microsomal uridine diphospho-glucuronosyltransferase (UDPGT): initial synthesis and characterization of an UDPGT immobilized enzyme reactor for the on-line study of glucuronidation.

The microsomal fraction of rat liver containing uridine diphospho-glucuronosyltransferase (UDPGT; EC 2.4.1.17) has been covalently immobilized on a high performance chromatographic support. In this study Nucleosil Si-500 silica was converted into diol-bonded silica and subsequently converted into an aldehyde form through oxidation with sodium periodate. The microsomal fraction was immobilized via Schiff base formation followed by reduction with sodium cyanoborohydride. The resulting immobilized enzyme reactor (IMER) was placed in a multi-dimensional chromatographic system which utilized a mixed mode (C18 and anion exchange) column to trap the parent compound and glucuronide and a C18 column to separate the substrate and product. The IMER system was used for the online glucuronidation of 4-methylumbelliferone (4Me7OHC) and acetaminophen (APAP). The Michaelis-Menten kinetic parameters (Km and Vmax) associated with the formation of 4Me7OHC and APAP glucuronides demonstrated that the immobilization had not significantly affected the enzymatic activity of the UDPGT relative to the non-immobilized enzyme. The IMER retained enzymatic activity for more than 6 weeks. The results of this study demonstrate an easy and convenient way to identify compounds which may be glucuronidated and to synthesize and characterize the resulting products.

Animals↗