Showing papers on "Bovine serum albumin published in 2002"
TL;DR: Fluorescence emission spectra of serum albumin in the presence of MMI or PTU, recorded at the excitation wavelengths 280 and 295 nm, clearly show that the studied drugs act as quenchers.
789 citations
TL;DR: This paper characterized nanoparticle uptake by endothelial cells in cell culture and demonstrated biocompatability of nanoparticles with the cells, demonstrating that nanoparticles could be used for localizing therapeutic agents or gene into endothelium.
509 citations
TL;DR: Characterizations and bioactivities show that the conjugate samples indeed contain both carbon nanotubes and BSA proteins and that the protein species are intimately associated with the nanot tubes.
Abstract: Carbon nanotubes are functionalized by bovine serum albumin (BSA) proteins via diimide-activated amidation under ambient conditions. The nanotube-BSA conjugates thus obtained are highly water-soluble, forming dark-colored aqueous solutions. Results from characterizations using atomic force microscopy (AFM), thermal gravimetric analysis, Raman, and gel electrophoresis show that the conjugate samples indeed contain both carbon nanotubes and BSA proteins and that the protein species are intimately associated with the nanotubes. Bioactivities of the nanotube-bound proteins are evaluated using the total protein micro-determination assay (the modified Lowry procedure). The results show that the overwhelming majority (∼90%) of the protein species in the nanotube-BSA conjugates remain bioactive.
507 citations
TL;DR: Caveolin-1 plays a dual regulatory role in controlling microvascular permeability: as a structural protein that is required for caveolae formation and caveolar transcytosis and as a tonic inhibitor of eNOS activity to negatively regulate the paracellular pathway.
319 citations
TL;DR: Polyamidoamine dendrimers interaction with bovine serum albumin was studied and it was shown that the generation 4 has a greater impact on spectral properties of serumalbumin than generation 3.5.
279 citations
TL;DR: This work has presented a review of the research on reactions of platinum(II) and (IV), gold(I) and(III), ruthenium(III) and rhodium( II) antitumoral compounds with serum albumin, and found that platinum and gold compounds are found to react with S-donors such as methionine and the Cys34 residues of albumin.
229 citations
TL;DR: Glycated albumin detected by the present method was significantly correlated with glycated album in detected by high-performance liquid-chromatographic (HPLC) method.
211 citations
TL;DR: In this article, Bovine serum albumin (BSA) was modified by covalent attachment of chlorogenic acid using different concentrations at pH 9. The structural changes were studied using circular dichroism, differential scanning calorimetry (DSC), intrinsic fluorescence, and binding of anilinonaphthalenesulfonic acid.
204 citations
TL;DR: It is the FFAs bound to albumin, rather than albumin itself, which cause severe tubulointerstitial damage by being reabsorbed into the proximal tubule, the first in vivo observation in which FFAs have caused severe Tubulointerstitial injury.
191 citations
TL;DR: In this article, the interactions between terazosin and bovine serum albumin (BSA) were studied by spectrofluorimetry, and the binding constants of BSA were measured at different temperatures.
176 citations
TL;DR: It is demonstrated that although increased water uptake induced by antacid excipients may improve the stability of the encapsulated proteins, the homogeneous acid neutralization effect is unique to antacidexcipients such as Mg(OH)2, which is necessary to maintain the Stability of proteins in acidic PLGA specimens.
TL;DR: The mechanism of aggregation of bovine serum albumin (BSA) by poly(allylamine) hydrochloride (PAH) is investigated as a function of the mixing ratio r defined as the ratio of the number of BSA molecules and PAH chains present in the solution, under pH conditions of strong binding between the two partners.
Abstract: The mechanism of aggregation of bovine serum albumin (BSA) by poly(allylamine) hydrochloride (PAH) is investigated as a function of the mixing ratio r defined as the ratio of the number of BSA molecules and PAH chains present in the solution, under pH conditions of strong binding between the two partners. It is found that as r increases the turbidity first increases, passes through a maximum at a value rmax before decreasing again. For small and large values of r, one forms small aggregates in the 10 nanometer size range, whereas at rmax, the size of the aggregates becomes of the order of micrometers. The structure of the aggregates appears to be independent of the history of the systems but depends only on the value of r despite the strong BSA/PAH binding. The desaggregation of the large aggregates formed at rmax by the addition of BSA or PAH is shown to be an isenthalpic process and is thus entirely entropically driven. Moreover, we prove that rmax corresponds to the state of the system where both the P...
TL;DR: The findings suggest that animal monoclonal antibodies specific for cow's milk proteins are able to recognize the major part of milk proteins from mammals bred in Mediterranean countries (sheep, goat, and buffalo); weak cross-reactivity was observed with milk proteins with mares and donkeys.
Abstract: Background Cross-reactivity between food allergens occurs when they share part of their amino acid sequence, or when their three-dimensional molecular structure causes them to have a similar capacity to bind specific antibodies. Objectives To review data from our laboratory on cross-reactivity between mammalian proteins (milk and meat allergens). Methods Studies used immunoelectrophoresis (sodium dodecyl sulfate-polyacrylamide gel electrophoresis/polyacrylamide gel electrophoresis and immunoblotting), and animal monoclonal antibodies. Results The findings suggest that animal monoclonal antibodies specific for cow's milk proteins are able to recognize the major part of milk proteins from mammals bred in Mediterranean countries (sheep, goat, and buffalo); weak cross-reactivity was observed with milk proteins from mares and donkeys. None of the antibodies used in our studies reacted with proteins from an exotic mammalian species: the camel. Similar cross-reactions were found with human circulating immunoglobulin E from children allergic to milk. With regard to beef allergy, monoclonal antibodies specific for bovine serum albumin cross-reacted only with ovine serum albumin, whereas the number of sera from allergic children able to recognize other mammalian serum albumins depended directly on the closeness of phylogenetic relationship between animal species and inversely on the percent identity with human serum albumin in the main epitopic sequence. Conclusion An area of heterogeneity between animal and human species in a critical amino acid sequence (epitope) of an allergen can determine the degree of immunogenic activity.
Patent•
08 Mar 2002
TL;DR: In this article, compositions comprising non-naturally occurring serum albumin binding moieties are described, together with methods of use thereof, e.g., for detecting or isolating serum bandit molecules in a solution, for blood circulation imaging, and for linking therapeutics or other molecules to albumin.
Abstract: Compositions comprising non-naturally occurring serum albumin binding moieties are described, together with methods of use thereof, e.g., for detecting or isolating serum albumin molecules in a solution, for blood circulation imaging, and for linking therapeutics or other molecules to albumin. Preferred serum albumin binding peptides having a high affinity for human serum albumin are particularly disclosed.
TL;DR: In this article, the effects of bovine serum albumin (BSA) on TNFα-induced expression of adhesion molecules in cultured human aortic endothelial cells (HAEC) were investigated.
Abstract: Objective: Leukocyte adhesion to, and transmigration across, the vascular endothelium are critical initiating steps in inflammation and atherosclerosis. We hypothesized that albumin, the major plasma protein, acts as an anti-inflammatory agent towards endothelial cells. Methods and Results: To test the hypothesis, we studied the effects of bovine serum albumin (BSA) on TNFα-induced expression of adhesion molecules in cultured human aortic endothelial cells (HAEC). We found that incubation of HAEC for 16 h with BSA (0.5–5%, w/v) dose-dependently inhibited TNFα-induced mRNA and protein expression of vascular cell adhesion molecule-1 (VCAM-1), but not intercellular adhesion molecule-1 nor E-selectin. Yeast recombinant human serum albumin exerted similar inhibitory effects on VCAM-1 expression, whereas γ-globulin was ineffective. BSA also significantly inhibited TNFα-induced adhesion of monocytic THP-1 cells to HAEC in a dose-dependent manner. Furthermore, BSA strongly inhibited activation and nuclear translocation of the transcription factor, nuclear factor-κB (NF-κB). For example, the physiologically relevant concentration of 5% BSA inhibited NF-κB activation by 90±7%, VCAM-1 mRNA and protein expression by 81±4 and 80±13%, respectively, and THP-1 adhesion by 73±9% ( n = 3). The inhibitory effect of BSA on TNFα-induced VCAM-1 expression was not attenuated by inhibition of intracellular GSH synthesis. Conclusions: Our data show that physiological concentrations of albumin selectively inhibit TNFα-induced upregulation of VCAM-1 expression and monocyte adhesion, most likely by inhibiting NF-κB activation in a GSH-independent manner.
TL;DR: In this article, surface-modified, polycarbonate track etched (PCTE) membranes were modified with monolayers of self-assembled thiols (HSC10H20COOH) on electroless gold.
Abstract: Control of external pH and ionic strength is used to separate proteins with surface-modified, nanoporous polycarbonate track etched (PCTE) membranes. The porous PCTE membranes were modified with monolayers of self-assembled thiols (HSC10H20COOH) on electroless gold. The hydraulic radius of the pores in the surface-modified membranes was 8.7 nm. Two proteins of nearly identical molecular weight, bovine serum albumin (BSA) and bovine hemoglobin (BHb), were used as the permeants. The fluxes of BSA and BHb through the membranes show maximum values at the isoelectric points (pI) of the proteins. At pH values above and below the pI, charge interactions between the proteins, their counterions, and the pore surface leads to a decrease in flux. The imposition of a difference in ionic strength across the membrane causes osmotic flow and leads to a significant increase in the protein fluxes and an enhancement of the selectivity of BSA over BHb. In protein separation experiments, the BSA and BHb fluxes are nearly 3 t...
TL;DR: High affinity binding of TCs with BSA signifies drug stability, however, excessive binding at higher TC concentrations in combination with Cu (II) induces conformational change in protein structure, which may exert detrimental effect on cellular protein.
TL;DR: The Cu(II) binding affinity of the N-terminal protein site is quantitatively higher when the single free thiol, Cys-34, is reduced (mercaptalbumin), compared to when it is oxidized or derivatized with N-ethylmaleimide, suggesting this increased affinity is due predominantly to entropic factors.
Abstract: The thermodynamics of Cu(II) and Ni(II) binding to bovine serum albumin (BSA) have been studied by isothermal titration calorimetry (ITC). The Cu(II) binding affinity of the N-terminal protein site is quantitatively higher when the single free thiol, Cys-34, is reduced (mercaptalbumin), compared to when it is oxidized or derivatized with N-ethylmaleimide. This increased affinity is due predominantly to entropic factors. At higher pH (approximately 9), when the protein is in the basic (B) form, a second Cu(II) binds with high affinity to albumin with reduced Cys-34. The Cu(II) coordination has been characterized by UV-vis absorption, CD, and EPR spectroscopy, and the spectral data are consistent with thiolate coordination to a tetragonal Cu(II), indicating this is a type 2 copper site with thiolate ligation. Nickel(II) binding to the N-terminal site of BSA is also modulated by the redox/ligation state of Cys-34, with higher Ni(II) affinity for mercaptalbumin, the predominant circulating form of the protein.
TL;DR: Several low‐abundance proteins of bovine colostrum and mature milk were identified using the immunoabsorption technique and two‐dimensional electrophoresis followed by microsequencing and mass spectrometry.
Abstract: We identified several low-abundance proteins of bovine colostrum and mature milk using the immunoabsorption technique and two-dimensional electrophoresis (2-DE) followed by microsequencing and mass spectrometry. Two major milk proteins, beta-casein and immunoglobulin G (IgG), were effectively removed from the milk using immunoabsorbents. Milk samples before and after immunoabsorption were separated by 2-DE. Protein identification of the spots on 2-DE was performed by either gel comparison, microsequencing, matrix-assisted laser desorption/ionization-time of flight mass-spectrometry (MALDI-TOF-MS), peptide mass fingerprinting or peptide sequencing using tandem MS by hybrid quadrupole/orthogonal acceleration time of flight-MS (Q-TOF). Significant differences in protein patterns were observed between the low-abundance proteins of colostrum and mature milk. In addition, several low-abundance proteins including fibrinogen beta-chain, chitinase 3-like 1, alpha-antitrypsin, complement C3 alpha-chain, gelsolin and apolipoprotein H were observed only in colostrum. However, the level of beta-casein fragments increased significantly during this lactation period. alpha-Lactalbumin and beta-lactoglobulin as well as some low-abundance proteins including bovine serum albumin, serotransferrin and lactoferrin were identified in both colostral and mature milk. Low-abundance proteins in bovine colostrum may have special physiologic relevance to the health and development of calves early in lactation.
TL;DR: The polyclonal antibody produced by immunization of rabbits with a conjugate of 4,4-bis (4-hydroxyphenyl) valeric acid and bovine serum albumin showed specific recognition of the bisphenol structure, while the cross reactions of other common phenolic compounds were lower than 1%.
TL;DR: The studies suggest that albumin has adequate binding capacity for the low plasma levels of V LCFA with 20 to 26 carbons, but the protein may not be able to bind longer chain VLCFA.
TL;DR: The results clearly demonstrate that nominal and relative toxic potencies of organochlorine pesticides and chlorophenols determined in vitro are substantially influenced by effects of protein binding on availability.
TL;DR: Immunohistochemistry of normal human brain demonstrated that IgM monoclonal antibody F95 stained central nervous system myelin and a subset of astrocytes, suggesting that certain astroglial filaments expressing glial fibrillary acidic protein also contain citrulline‐bearing components.
Abstract: Using hybridoma technology, an IgM monoclonal antibody (mAb), designated as F95, was developed against a deca-citrullinated peptide (DCP) consisting of 10 citrulline residues and a carboxyl Gly-Gly-Cys through which DCP was covalently linked to an activated carrier protein, keyhole limpet hemocyanin (KLH). Clones were selected on the basis of not reacting with human unmodified and noncitrullinated myelin basic protein (MBP), MBP-C1, but reacting well with human citrullinated MBP (MBP-C8). When tested by ELISA, this mAb demonstrated minimal reactivity with human MBP-C1, varying reactivity with the C2-C5 isomers of human MBP, moderate binding with guinea pig MBP-C8, and strong reactivity with human MBP-C8. By ELISA, mAb F95 was directed predominantly against citrulline, not MBP, as revealed by its binding to DCP linked with activated KLH, bovine serum albumin (BSA), or ovalbumin (OA), but not with KLH, BSA, or OA alone. Immunohistochemistry of normal human brain demonstrated that F95 stained central nervous system myelin and a subset of astrocytes. Given the citrulline-directed features of mAb F95, this immunohistochemical pattern suggests that certain astroglial filaments expressing glial fibrillary acidic protein also contain citrulline-bearing components. These potentially implicate citrullinated proteins, notably in astroglial filaments, in a variety of normal and pathological neurobiological processes.
TL;DR: It was found that while qualitative aspects of interaction could be reproduced, the quantitative parameters could not, and a very strong influence of phosphate ions on the kinetics of Ni(II) interaction was found.
Abstract: A comparative study of thermodynamic and kinetic aspects of Cu(II) and Ni(II) binding at the N-terminal binding site of human and bovine serum albumins (HSA and BSA, respectively) and short peptide analogues was performed using potentiometry and spectroscopic techniques. It was found that while qualitative aspects of interaction (spectra and structures of complexes, order of reactions) could be reproduced, the quantitative parameters (stability and rate constants) could not. The N-terminal site in HSA is much more similar to BSA than to short peptides reproducing the HSA sequence. A very strong influence of phosphate ions on the kinetics of Ni(II) interaction was found. This study demonstrates the limitations of short peptide modelling of Cu(II) and Ni(II) transport by albumins.
TL;DR: Several clinically important proteins and enzymes have been immobilised onto fine magnetic particles using carbodiimide as a coupling agent and the possible applications in biomedicine and biotechnology are discussed.
Abstract: Several clinically important proteins and enzymes (bovine serum albumin (BSA), glucose oxidase (GOD) (EC 1.1.3.4), streptokinase (EC 3.4.99.0), chymotrypsin (EC 3.4.21.1) and dispase (EC 3.4.24.3)), respectively, have been immobilised onto fine magnetic particles using carbodiimide as a coupling agent. The coupling reactions of these substances were carried out using various ratios of magnetic particles to protein, and different values of pH to determine the optimum conditions of immobilisation. The possible applications in biomedicine and biotechnology of this method of immobilisation are discussed.
TL;DR: The present study has provided useful basic information for a therapeutic strategy and the molecular design of succinylated proteins for use as drug carriers and therapeutic agents per se for SR-mediated targeting in vivo.
Abstract: In vivo disposition characteristics of succinylated (Suc-) proteins were studied after intravenous injection in mice in relation to their molecular characteristics as negatively charged macromolecules Recombinant superoxide dismutase (SOD; molecular mass, 32 kDa), bovine serum albumin (BSA; molecular mass, 67 kDa), and bovine IgG (molecular mass, 150 kDa) were used to produce succinylated derivatives with different degrees of modification 111In-labeled Suc-SODs were rapidly excreted into the urine with no significant hepatic uptake In contrast, 111In-Suc-BSA and Suc-IgG were significantly taken up by liver nonparenchymal cells via scavenger receptors (SRs) according to the degree of succinylation and the dose injected Interestingly, highly succinylated BSAs exhibited significant accumulation in the kidney at higher doses when the hepatic uptake was saturated Pharmacokinetic analysis demonstrated that the hepatic uptake of succinylated proteins depended on the molecular size and the estimated surface density of succinylated amino residues Further analysis based on a physiological pharmacokinetic model, involving a saturable process with Michaelis-Menten kinetics, revealed that the surface density of negative charges was correlated with the affinity of larger succinylated proteins for the hepatic SRs Thus, the present study has provided useful basic information for a therapeutic strategy and the molecular design of succinylated proteins for use as drug carriers and therapeutic agents per se for SR-mediated targeting in vivo
TL;DR: Electrostatic forces play a major role in the interaction between the oligonucleotide and the nanoparticles, and only the olig onucleotide incorporated into the nanoparticle matrix was protected against enzymatic degradation.
TL;DR: In this paper, three new benzazole isothiocyanate (BzITC) fluorescent dyes, 2-(5′-isothiocoyanate-2′-hydroxyphenyl)benzoxazole, 2.5-b]pyridine and 2.7-b]-benzimidazole were synthesized, purified until optical purity grade and characterised by elemental analysis, 1 H NMR, IR, UV-VIS and fluorescence spectroscopy.
Abstract: Three new benzazole isothiocyanate (BzITC) fluorescent dyes, 2-(5′-isothiocyanate-2′-hydroxyphenyl)benzoxazole, 2-(5′-isothiocyanate-2′-hydroxyphenyl)benzimidazole and 2-(5′-isothiocyanate-2′-hydroxyphenyl)oxazole[4,5-b]pyridine, were synthesised, purified until optical purity grade and characterised by elemental analysis, 1 H NMR , IR, UV–VIS and fluorescence spectroscopy. These dyes exhibit an intense fluorescence emission with a large Stokes shift due to an excited state intramolecular proton transfer (ESIPT) mechanism. The BzITCs were also studied for labelling three proteins (bovine serum albumin (BSA), concanavalin-A (con-A) and rabbit immunoglobulin G (rabbit IgG)) and the resulting conjugates presented good and stable fluorescence. A simple assay for detection of these proteins was reported here. The method is based on the direct fluorescence detection of protein-labelled with BzITC fluorophores after polyacrylamide gel electrophoresis and present potential use as fluorescent probes for proteins.
TL;DR: The data suggest that thiolation of the protein-bound acrolein may be involved in redox alteration under oxidative stress, whereby oxidative stress generates the increased production of ac rolein and its protein adducts that further potentiate oxidative stress via the depletion of GSH in the cells.
TL;DR: The size of the guest molecule is an important parameter when electrically-stimulated drug release is desired, but further work obviously needs to be carried out with a larger range of molecular weights and conformations.