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Showing papers on "Human serum albumin published in 1985"


Journal ArticleDOI
TL;DR: The results confirm the concept that although DBP is the principal protein carrier of 1,25-(OH)2D in serum, albumin is a major secondary carrier, especially in patients with low DBP levels.
Abstract: Using the technique of centrifugal ultrafiltration isodialysis to measure the free concentration of 1,25-dihydroxyvitamin D [1,25-(OH)2D], we determined the affinity of serum proteins for 1,25-(OH)2D both by Scatchard analysis (increasing ligand concentration at fixed binding site concentrations) and by a novel analysis in which the binding site concentrations were varied (serial dilution) at fixed ligand concentrations. The high affinity binding constant in serum for 1,25-(OH)2D was 3.7 X 10(7) M-1 by Scatchard analysis and 4.2 X 10(7) M-1 by serial dilution analysis. Human serum albumin had a much lower affinity for 1,25-(OH)2D (5.4 X 10(4) M-1). When vitamin D-binding protein (DBP) was selectively removed from serum by an actin affinity column, the affinity of the remaining serum proteins for 1,25-(OH)2D was that of albumin. Postulating a two-site model (DBP and albumin) for transport of 1,25-(OH)2D in serum and incorporating the estimated affinity constants of DBP and albumin for this metabolite, we calculated that 85% of total circulating 1,25-(OH)2D is transported in blood bound to DBP in normal individuals (0.4% is free and 14.6% is bound to albumin). In patients with liver disease, 73% is bound to DBP (1.1% is free and 25.9% is bound to albumin). Using this same two site model, we found a reasonable correlation (r = 0.612; P less than 0.001) between the measured free 1,25-(OH)2D level and the calculated free 1,25-(OH)2D level in serum based on albumin and DBP concentrations in 16 normal subjects and 16 patients with liver disease. These results confirm the concept that although DBP is the principal protein carrier of 1,25-(OH)2D in serum, albumin is a major secondary carrier, especially in patients with low DBP levels.

247 citations


Journal ArticleDOI
TL;DR: It appears that anti-B.
Abstract: The objective of the present study was to determine the relationship between concentrations of antibodies in serum and those in gingival crevicular fluid (GCF) of patients with juvenile periodontitis and severe periodontitis. Most antigens used to quantitate antibodies were obtained from a panel of bacteria associated with juvenile periodontitis or severe periodontitis. We further investigated variation in antibody titer among different periodontal sites and the extent to which antibody in GCF is locally derived. Titers of antibody, total immunoglobulin G (IgG), and human serum albumin were determined with sensitive radioimmunoassays. The relationship between serum and GCF antibody was complex. Both person-to-person variability and marked variability within the same subject were found among different sites of similar clinical status. The site-to-site variability was found not only for antibody reactive with periodontal organisms, but also for antitetanus toxoid, total IgG, and even human serum albumin. Generally the variability was in the degree of depression of the level in GCF relative to that in serum. However, anti-Bacteroides gingivalis and anti-Actinobacillus actinomycetemcomitans in GCF often exceeded the level in serum. When antibody titers in serum and GCF were calculated per milligram of human serum albumin, most of the apparent depressions of antibody in GCF disappeared. The ratio of antibody in serum to that in GCF approached unity for all organisms except B. gingivalis and A. actinomycetemcomitans Y4, which were markedly elevated. Furthermore, the level of IgG per milligram of human serum albumin in GCF was about twice the level in serum. We believe that human serum albumin reflects serum contribution to the GCF, and we therefore attribute the increased level of IgG per milligram of albumin in GCF to local synthesis. It appears that anti-B. gingivalis and anti-A. actinomycetemcomitans represent an important portion of this local antibody synthesis, since most seropositive patients with severe or juvenile periodontitis had at least one site elevated, and the magnitudes of the elevations were large in many sites. Those sites yielding elevated antibody exhibited no obvious differences in clinical parameters of probeable depth or attachment level as compared with sites in which antibody levels in GCF were similar to serum levels. Elevated antibody in GCF may relate to changes in disease activity that are not detectable by usual clinical measures.

162 citations


Journal ArticleDOI
TL;DR: Comparisons of equilibrium dialysis, ultrafiltration, and ultracentrifugation were compared to determine their reliability and applicability in the study of binding of an anticonvulsant drug, valproic acid, by plasma proteins.
Abstract: Equilibrium dialysis, ultrafiltration, and ultracentrifugation were compared to determine their reliability and applicability in the study of binding of an anticonvulsant drug, valproic acid, by plasma proteins. We studied drug binding with pooled serum and with solutions of human serum albumin at physiological concentrations. We compared binding characteristics such as number of binding sites, affinity constants, and percent of binding as measured by each method in the therapeutic range for valproic acid. Results by ultracentrifugation differed from those by equilibrium dialysis and ultrafiltration, which agreed reasonably well with each other.

140 citations


Journal ArticleDOI
TL;DR: A masticatory-lubrication assay system is reported for the first time to assess the lubricating properties of salivary constituents and it is demonstrated that deglycosylation of the PRG altered the nature of its interaction with albumin.
Abstract: We report for the first time a masticatory-lubrication assay system to assess the lubricating properties of salivary constituents. The lubricating ability of the proline-rich glycoprotein (PRG) of parotid saliva was enhanced by human serum albumin. The interactive effect of albumin was abolished by chemically deglycosylating the glycoprotein. Fluorescence spectroscopy with a hydrophobic probe verified the existence of a PRG-albumin complex and demonstrated that deglycosylation of the PRG altered the nature of its interaction with albumin.

140 citations


Journal ArticleDOI
TL;DR: This labeling technique can be used in the preparation of intravascular NMR contrast agents (like paramagnetically-labeled human serum albumin) or target-specific agents (labeled monoclonal antibodies or fibrinogen).

127 citations


Journal ArticleDOI
TL;DR: The present findings support the proposal of four separate primary binding sites for warfarin, digitoxin (and salicylate), diazepam and Phenol Red.
Abstract: Binding of warfarin, digitoxin, diazepam, salicylate and Phenol Red, individually or in different pair combinations, to defatted human serum albumin at ligand/protein molar ratios less than 1:1 was studied at pH 7.0. The binding was determined by ultrafiltration. Some of the experiments were repeated with the use of equilibrium dialysis in order to strengthen the results. Irrespective of the method used, all ligands bind to one high-affinity binding site with an association constant in the range 10(4)-10(6) M-1. High-affinity binding of the following pair of ligands took place independently: warfarin-Phenol Red, warfarin-diazepam, warfarin-digitoxin and digitoxin-diazepam. Simultaneous binding of warfarin and salicylate led to a mutual decrease in binding of one another, as did simultaneous binding of digitoxin and Phenol Red. Both effects could be accounted for by a coupling constant. The coupling constant is the factor by which the primary association constants are affected; in these examples of anti-co-operativity the factor has a value between 0 and 1. In the first example it was calculated to be 0.8 and in the latter 0.5. Finally, digitoxin and salicylate were found to compete for a common high-affinity binding site. The present findings support the proposal of four separate primary binding sites for warfarin, digitoxin (and salicylate), diazepam and Phenol Red. An attempt to correlate this partial binding model for serum albumin with other models in the literature is made.

105 citations


Journal ArticleDOI
TL;DR: A specific immune response was defined for each compound belonging to the same metabolic pathway, and the cross‐reactivity ratios were found to be smallest for the most immunoreactive conjugates.
Abstract: Antisera were raised against tryptophan, 5-hydroxytryptophan, 5-hydroxytryptamine, 5-methoxytryptophan, and 5-methoxytryptamine, by conjugating each molecule to bovine serum albumin and to human serum albumin via glutaraldehyde, in such a way as to preserve the original part. Antibody specificity was tested with the enzyme-linked immunosorbent assay method. The specificity of each anti-indolealkylamine-glutaraldehyde antibody was established with competition experiments by using an adsorbed immunogenic conjugate and indolealkylamines either free or conjugated with poly-L-lysine. The nonconjugated compounds were poorly recognized. In the same way, the nonreduced conjugates always appeared less immunoreactive than the reduced ones. Calculated from the specificity study of each antiserum, the cross-reactivity ratios were found to be smallest for the most immunoreactive conjugates. Thus, a specific immune response was defined for each compound belonging to the same metabolic pathway.

91 citations


Journal Article
TL;DR: IgG antibodies were found to increase the uptake of circulating dinintrophenylated human serum albumin preparations by the nonparenchymal liver cells in rats, implying that serum complement and hepatic C3 receptors are essential for the physiological clearance of circulating immune complexes.
Abstract: IgG antibodies were found to increase the uptake of circulating dinintrophenylated human serum albumin (DNP-HSA) preparations by the nonparenchymal liver cells in rats. Highly DNP-conjugated HSA was taken up by the Kupffer cells both when given alone and when complexed by IgG. More lightly DNP-conjugated HSA was taken up mainly by the liver endothelial cells. Here, IgG promoted the antigen uptake both by the Kupffer cells and by the endothelial cells. Uptake of IgG immune complexes (IgG-ICs) by the sinusoidal endothelial cells of the liver is a new aspect on the function of these cells. Whether or not this phenomenon is Fc receptor-mediated is discussed. A heat-labile serum factor was found to direct the ICs to the Kupffer cells. This implies that serum complement and hepatic C3 receptors are essential for the physiological clearance of circulating immune complexes.

88 citations


Journal Article
TL;DR: The receptor for polymerized human as well as chimpanzee serum albumins has been identified on the 55-amino acid polypeptide coded by the pre-S region of hepatitis B virus DNA.
Abstract: The receptor for polymerized human as well as chimpanzee serum albumins has been identified on the 55-amino acid polypeptide coded by the pre-S region of hepatitis B virus DNA. Monoclonal antibodies were raised against a synthetic polypeptide of 19 amino acid residues representing a hydrophilic region of the pre-S amino acid sequence deduced from hepatitis B virus DNA. Sheep erythrocytes fixed with glutaraldehyde were coated with monoclonal antibody against the synthetic polypeptide to develop a hemagglutination assay for pre-S polypeptide. The pre-S polypeptide was detected in the serum containing hepatitis B surface antigen particles along with hepatitis B e antigen, with titers in parallel with those of the receptor for polymerized human serum albumin.

75 citations


Journal ArticleDOI
TL;DR: High-performance liquid chromatographic analysis of human serum albumin showed three peaks, the principal component corresponding to human mercaptalbumin (HMA) and the secondary and tertiary components to nonmercaptalbumIn (HNA), indicating that HMA is a covalent carrier protein for sulphur-containing amino acids.

74 citations


Journal ArticleDOI
TL;DR: In this article, microspheres intended for intra-arterial targeting of cytotoxic agents have been prepared from human serum albumin using a water-in-oil emulsion technique with chemical cross-linking of the protein.

Journal ArticleDOI
TL;DR: In this paper, the applicability of a model of specific and non-specific binding sites for the binding of organic ligands to serum albumin was examined statistically, showing that the model fitted the data for binding of all compounds very well.

Journal ArticleDOI
TL;DR: The adsorbent glycinehydroxamate-Sepharose 6B, charged with Fe3+ under specified conditions, is reported, and was used at various pH values for chromatography of the following proteins: lysozyme, cytochrome c, avidin, bovine pancreatic RNase, myoglobin, ovalbumin and human serum albumin.

Journal ArticleDOI
TL;DR: The binding of diclofenac to human serum albumin (HSA) and to lipoproteins was studied in vitro by equilibrium dialysis and the evidence of two specific binding sites on HSA was confirmed by circular dichroism data.

Journal ArticleDOI
TL;DR: The kinetics of quenching of tryptophan fluorescence in HSA by PII indicates that there is one main porphyrin-binding site affecting this fluorescence, which seems to have a slightly higher affinity for PII than the remaining sites.

Patent
28 Mar 1985
TL;DR: In this paper, an interleukin-2 composition which comprises human serum albumin, a reducing compound or a combination thereof is adjusted as showing pH of 3 to 6 as a solution.
Abstract: The present invention provides an interleukin-2 composition which comprises human serum albumin, a reducing compound or a combination thereof and is adjusted as showing pH of 3 to 6 as a solution. The composition of the present invention is characterised in that the interleukin-2 activity is lost little during storage and in the process of freezing and lyophilization

Journal ArticleDOI
TL;DR: Two models are proposed for the protein-binding of porphyrin monomers and dimers in a porphyrs system having both species: a competitive model, where each protein molecule has only one binding site, which can be occupied by either a monomer or a dimer; a non-competitive model, which has two binding sites, one for monomer and one for dimers.
Abstract: The binding equilibrium of deuteroporphyrin IX to human serum albumin and to bovine serum albumin was studied, by monitoring protein-induced changes in the porphyrin fluorescence and taking into consideration the self-aggregation of the porphyrin. To have control over the latter, the range of porphyrin concentrations was chosen to maker dimers (non-covalent) the dominant aggregate. Each protein was found to have one high-affinity site for deuteroporphyrin IX monomers, the magnitudes of the equilibrium binding constants (25 degrees C, neutral pH, phosphate-buffered saline) being 4.5 (+/- 1.5) X 10(7) M-1 and 1.7 (+/- 0.2) X 10(6) M-1 for human serum albumin and for bovine serum albumin respectively. Deuteroporphyrin IX dimers were found to bind directly to the protein, each protein binding one dimer, with high affinity. Two models are proposed for the protein-binding of porphyrin monomers and dimers in a porphyrin system having both species: a competitive model, where each protein molecule has only one binding site, which can be occupied by either a monomer or a dimer; a non-competitive model, where each protein molecule has two binding sites, one for monomers and one for dimers. On testing the fit of the data to the models, an argument can be made to favour the non-competitive model, the equilibrium binding constants of the dimers, for the non-competitive model (25 degrees C, neutral pH, phosphate-buffered saline), being: 8.0 (+/- 1.8) X 10(8) M-1 and 1.2 (+/- 0.6) X 10(7) M-1 for human serum albumin and bovine serum albumin respectively.

Journal ArticleDOI
TL;DR: An association between the presence of IgE or total antibody to ETO-HSA and immediate anaphylactic reactions in this group of 65 patients receiving hemodialysis is clearly demonstrated.
Abstract: We have measured total antibody and IgE directed against ethylene oxide-altered human serum albumin (ETO-HSA) in the sera of 24 patients who have experienced anaphylaxis during hemodialysis and of 41 patients who have not had such episodes during hemodialysis. ETO is used to sterilize dialyzers and other medical equipment. The geometric mean level of IgE to ETO-HSA in patients with reactions (0.9 ng ETO-HSA bound to IgE per milliliter of serum) is significantly higher than in nonreacting patients (0.1 ng/ml, p p p p

Journal Article
TL;DR: These studies support the earlier experiments which demonstrated that C5b-9 is in a different molecular configuration on the surface of serum-resistant GC from that on thesurface of Serum-sensitive GC or resistant GC rendered sensitive with bactericidal antibody.
Abstract: In this study, we examined the bacterial constituents associated with the complement C5b-9 complex in detergent extracts from serum-treated Neisseria gonorrhoeae (GC). 125I surface-labeled GC were incubated in 10% serum, were washed, and were solubilized in the zwitterionic sulfobetaine detergent SB12. Immunoprecipitation of 125I-GC from the extract with anti-C5 Sepharose was followed by 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiography of immunoprecipitated material. Polyacrylamide gel analysis of surface-labeled 125I-GC showed prominent bands for proteins I and III for both serum-resistant GC strain 6305 and serum-sensitive GC strain 7189. These same bands were visible with similar intensity in the SB12 extracts from presensitized and non-presensitized 6305 and 7189 after serum incubation. For those organisms bearing bactericidal C5b-9 (6305 + IgG and 7189 +/- IgG), additional distinctive bands immunoprecipitated with antibody to C5 Sepharose. These components were of 93,000, 44,000 40,000, and 15,000 daltons for 6305 + IgG, and were of 90,000, 50,000, 44,000, and 19,000 daltons for 7189 +/- IgG. Nonbactericidal C5b-9 extracted from the surface of 6305 incubated in serum, but not sensitized with antibody, was not associated with these distinctive proteins. However, this nonbactericidal C5b-9 did have a different pattern of associated bacterial surface constituents from that observed in control samples incubated with antibody to human serum albumin, which were similar to those with nonserum-incubated organisms. These studies support our earlier experiments which demonstrated that C5b-9 is in a different molecular configuration on the surface of serum-resistant GC from that on the surface of serum-sensitive GC or resistant GC rendered sensitive with bactericidal antibody.

Patent
13 Sep 1985
TL;DR: An improved process for recovering and purifying lipophilic recombinant proteins such as human β-interferon and interleukin-2 from their hosts yields a protein preparation which may be formulated into a stable pharmaceutical composition having a therapeutically effective amount of the biologically active recombinant lipid protein dissolved in a non-toxic, inert, therapeutic compatible aqueous-based carrier medium at a pH of 6.8 to 7.8 which medium also contains a stabilizer for the protein, such as normal serum albumin, normal albumin and human
Abstract: An improved process for recovering and purifying lipophilic recombinant proteins such as human β-interferon and interleukin-2 from their hosts yields a protein preparation which may be formulated into a stable pharmaceutical composition having a therapeutically effective amount of the biologically active recombinant lipophilic protein dissolved in a non-toxic, inert, therapeutically compatible aqueous-based carrier medium at a pH of 6.8 to 7.8 which medium also contains a stabilizer for the protein, such as human serum albumin, normal serum albumin and human plasma protein fraction.

Journal Article
TL;DR: All observations were consistent with a binding process involving albumin and the warfarin anion, without participation of hydrogen ions and not influenced by the N-B conformational transition of albumin.
Abstract: Reversible binding of warfarin to defatted serum albumin was studied by equilibrium dialysis at pH 7.4, in a 66 mM sodium phosphate buffer at 37 degrees. The binding isotherm could be described by two stoichiometric binding constants, K1 in the range 141,000 to 192,000 M-1 and K2 at 39,000 to 57,000 M-1. At least two additional molecules could be bound but gave indeterminate binding constants. The product K3 X K4 was about 4.7 X 10(7) M-2. Different site models were possible, either one high affinity and several low affinity sites, or two high affinity sites, cooperative, independent, or anticooperative, together with two low affinity sites. Binding affinity for the first warfarin molecule did not vary with pH in the interval from 6 to 9. The affinity decreased with increasing concentrations of sodium sulfate, sodium chloride, and calcium chloride, depending upon ionic strength. Specific effects of chloride and calcium ions were not observed. Light absorption spectra indicated that the warfarin anion was bound to albumin. All observations were consistent with a binding process involving albumin and the warfarin anion, without participation of hydrogen ions and not influenced by the N-B conformational transition of albumin.

Journal ArticleDOI
TL;DR: The results indicate that the equine estrogens bind to SHBG and albumin in a manner similar to that of E1 and E2, and that the low MCR of EqS reported previously may be due to its binding to albumin.
Abstract: The binding of ring B unsaturated equine estrogens, equilin sulfate (EqS), equilin (Eq), and 17 beta-dihydroequilin (17 beta-Eq) with human serum proteins was determined and compared with the binding of estrone sulfate (E1S), estrone (E1), estradiol (E2), testosterone (T), and 5 alpha-dihydrotestosterone (5 alpha-DHT). Undiluted serum or 5% human serum albumin (HSA) was incubated with 3H-labeled steroids at 37 C, then subjected to gel filtration at 4 C. Gel filtration of serum from Premarin-treated postmenopausal women or normal women incubated with Eq, E1, E2, or 5 alpha-DHT showed two peaks of radioactivity associated with proteins with average apparent mol wt of 128,000 and 68,000 and average Stokes radii of 48.6 and 34.9 A. These values correspond to those reported for sex hormone-binding globulin (SHBG) and albumin, respectively. Binding to SHBG and albumin was confirmed by removing SHBG or albumin from the serum with Concanavalin-A Sepharose 4B gel or CM-Affi Gel Blue, respectively. In the case of [3H]EqS and [3H]E1S, binding to SHBG was not detectable, and only a peak of radioactivity associated with albumin was found. However, under these conditions, the binding of estrogens to SHBG in serum from normal men was not detectable. Incubation of the above steroids with 5% HSA followed by gel filtration resulted in a single peak of radioactivity associated with the protein peak. Using ultrafiltration dialysis followed by Scatchard analysis, at least two sets of binding sites were found for the interaction of HSA with EqS or E1S. The high and low affinity binding sites had association constants k1 and k2 of approximately 0.9-1.1 (X 10(5) M-1) and 0.5-0.8 (X 10(4) M-1). In contrast with Eq and E1, only the low affinity binding sites were found (apparent Ka congruent to 1 X 10(4) M-1). The binding constants of some estrogens and androgens to SHBG at 37 C determined by competitive Scatchard analysis using DEAE filter assay and [3H]5 alpha-DHT were 0.15, 0.07, 0.22, 0.29, 2.70, and 4.53 (X 10(9) M-1) for Eq, E1, 17 beta-Eq, E2, T, and 5 alpha-DHT, respectively. These results indicate that the equine estrogens bind to SHBG and albumin in a manner similar to that of E1 and E2, and that the low MCR of EqS reported previously may be due to its binding to albumin.

Journal ArticleDOI
TL;DR: Electron microscopy investigations on cell damages caused by loading with liposome-bound porphyrins and subsequent illumination show that the plasmatic membrane is one important cell site of p Morphyrin interaction and photodynamic effect.

Book ChapterDOI
TL;DR: It is shown that porphyrin have a marked affinity for tissues with a high lipoprotein content, and attention has been drawn to hemopexin, a protein known to carry heme into hepatocytes, and to lipoproteins.
Abstract: It is of importance to determine how porphyrins are transported in blood, for two main reasons: (i) porphyrins are used for fluorescence detection and photodynamic therapy of tumors, and (ii) certain diseases such as the porphyrias and disorders such as lead intoxiation lead to an increased level of porphyrins in the blood. Until recently, albumin was thought to be the main porphyrin binding protein in serum. However, since most porphyrins are cleared through the liver, and since albumin is not taken up by hepatocytes, it has bejn conjectured that other porphyrin-binding elements exist in serum1. Attention has been drawn to hemopexin, a protein known to carry heme into hepatocytes, and to lipoproteins. Already in 1956, it was shown that porphyrin have a marked affinity for tissues with a high lipoprotein content 2.

Patent
08 Apr 1985
TL;DR: In this paper, an interleukin-2 composition which comprises human serum albumin, a reducing compound or a combination thereof is adjusted as showing pH of 3 to 6 as a solution.
Abstract: The present invention provides an interleukin-2 composition which comprises human serum albumin, a reducing compound or a combination thereof and is adjusted as showing pH of 3 to 6 as a solution. The composition of the present invention is characterized in that interleukin-2 activity is minimized during storage and in the processes of freezing and lyophilization.

Journal ArticleDOI
TL;DR: It is demonstrated that the enhanced albumin passage through the wall of the microvasculature characteristically found in long-term Type 1 diabetic patients with clinical microangiopathy is pressure-dependent to a large extent.
Abstract: The effect of acute arterial blood pressure lowering upon albumin extravasation was studied in 10 patients with nephropathy and retinopathy due to long-standing Type 1 (insulin-dependent) diabetes. The following variables were measured: transcapillary escape rate of albumin (initial disappearance of intravenously injected 125I-labelled human serum albumin), and urinary albumin excretion rate (radial immuno-diffusion). The study was performed twice within 2 weeks, with the patients receiving an intravenous injection of either clonidine (225 μg) or saline (0.154 mmol/l). The clonidine injection induced the following changes: arterial blood pressure decreased from 134/87 to 107/73 mmHg (p<0.01), transcapillary escape rate of albumin declined from 8.1 to 6.7% of the intravascular mass of albumin/h (p<0.01), albuminuria diminished from 1434 to 815 μg/min (p<0.01), and plasma volume raised slightly from 2916 to 2995ml (p<0.05). Our findings demonstrate that the enhanced albumin passage through the wall of the microvasculature characteristically found in long-term Type 1 diabetic patients with clinical microangiopathy is pressure-dependent to a large extent. This may be due to elevated hydrostatic pressure in the microcirculation.

Journal ArticleDOI
TL;DR: In this paper, contact angle determinations with two liquids, on hydrated as well as on dried protein layers, the long-range and short-range contributions to the protein surface tensions were determined.
Abstract: By means of contact angle determinations with two liquids, on hydrated as well as on dried protein layers, the long-range and the short-range contributions to the protein surface tensions, and from these the protein (ΔG 131) and the protein-ligand (ΔG 132) free energies of interaction in aqueous media, were determined. For human serum albumin (HSA), human IgG, and human IgA, the differences between ΔG 131 HYDRATED and ΔG 131 DRY were connected with the behavior of these proteins in low concentrations of (NH4)2SO4 versus saturated (NH4)2SO4 solutions. By interpolation, intermediate points are found that correlate well with the known salting-out properties of these three proteins. On the basis of the data, it is predicted that the precipitation of IgG by 1/3 saturated (NH4)2SO4 is preventable, or reversible, by the admixture of 15% dimethylsulfoxide; both predictions are confirmed experimentally. From the ΔG 132 values found, it is shown that HSA and IgG should attach to phenyl ligands under physiological conditions, but that IgA is so hydrophilic that it only can adhere to phenyl ligands after partial dehydration brought about when admixed with 1 M (NH4)2SO4. Closer analysis of the values obtained for the long-range and short-range components of the surface tensions of HSA, IgG, and IgA allow deeper insight into their functional, chemical, and physicochemical properties.

Journal ArticleDOI
TL;DR: The method involving alkaline benzethonium chloride with measurement at 450 nm had the best sensitivity within the range of linearity and the most consistent bias among the three globulin fractions, defining the dilemma for valid calibration of these methods for total serum protein in cerebrospinal fluid and urine.
Abstract: We used human serum protein fractions to evaluate the sensitivity and bias of three turbidimetric methods for determining concentrations of proteins. Each fraction (Cohn Fractions II, III, IV, and V) was assigned a protein concentration value that was determined by the biuret method, which we calibrated with purified monomer of human serum albumin. All three turbidimetric methods (those involving sulfosalicylic acid/sodium sulfate, trichloroacetic acid, and alkaline benzethonium chloride) gave acceptable results for Fraction V with crystallized human serum albumin as the reference material, but there was bias by each of the three methods for the three globulin fractions. The method involving alkaline benzethonium chloride with measurement at 450 nm had the best sensitivity within the range of linearity and the most consistent bias among the three globulin fractions. These results define the dilemma for valid calibration of these methods for total serum protein in cerebrospinal fluid and urine.

Journal ArticleDOI
TL;DR: An apparent increase in the molecular weight of ETO-HSA as compared to HSA suggests that antibody is directed against a new antigenic determinant on the Eto-protein moiety, probably an ETO -amino acid determinant like lysine.
Abstract: Anaphylactic reactions to dialysis have been associated with antibody to human serum albumin (HSA) that has been reacted with ethylene oxide (ETO). ETO, an agent used to sterilize dialyzers, is a potent alkylator of proteins. We have studied the antigenic characteristics of ETO-HSA. By use of immunoelectrophoresis we have demonstrated differences in electrophoretic mobility between HSA and ETO-HSA. By gel chromatography we have observed an apparent increase in the molecular weight of ETO-HSA as compared to HSA. Antigenic activity of ETO-HSA as determined by the technique of Lidd and Farr is found in all molecular weight fractions. Cutaneous reactivity to ETO-HSA can be transferred to subhuman primates. That reactivity is lost if the sera are heated to 56 °C for 4 hours. Antibody activity to ETO-HSA cannot be inhibited by ETO-glycine or HSA but can partially be inhibited by ETO-polylysine or ETO-ovalbumin. This suggests that antibody is directed against a new antigenic determinant on the ETO-protein moiety, probably an ETO-amino acid determinant like lysine.

Journal ArticleDOI
TL;DR: The percentage of oxazepam bound to plasma proteins in patients with renal impairment was lower than in normal volunteers, and this lower binding can neither be attributed to lower albumin concentrations because of the large binding capacity of the protein and linearity of N X K nor to displacement by elevated concentrations of glucuronide conjugates, but it may be ascribed partly to increased plasma fatty acids.