scholarly journals Trichloressigsäure-Aceton-Extraktion von Albuminen aus Seren und Antigen-Antikörper-Präzipitaten

1957 ◽  
Vol 12 (12) ◽  
pp. 777-783 ◽  
Author(s):  
Ekkehard Kallee ◽  
Friedrich Lohss ◽  
Willmar Oppermann
Keyword(s):  
Good Yield ◽  
Trichloroacetic Acid ◽  
Human Albumin ◽  
Binding Ability ◽  
Dye Binding

1. Highly purified albumin fractions can be obtained by precipitating human, rabbit, or rat sera with trichloroacetic acid, and extracting the precipitates with acetone.2. Human albumin thus prepared is identical with the original albumin in its electrophoretic properties, its dye-binding ability, and in its immunological behavior.3. After precipitating human albumin with antibodies the albumin antigen can be extracted in a good yield by use of trichloroacetic acid and acetone without losing the electrophoretic or antigenic characteristics of albumin.

scholarly journals STUDIES ON EXPERIMENTAL HYPERTENSION

1943 ◽  
Vol 78 (1) ◽  
pp. 67-74 ◽  
Author(s):  
Yale J. Katz ◽  
Harry Goldblatt
Keyword(s):  
Blood Pressure ◽  
Sodium Hydroxide ◽  
Ammonium Sulfate ◽  
Good Yield ◽  
Trichloroacetic Acid ◽  
Experimental Hypertension ◽  
Distilled Water ◽  
Successive Treatment ◽  
Physiological Properties ◽  
Hog Kidney

1. Extraction of finely ground fresh hog kidney with distilled water adjusted to pH 7.8 with sodium hydroxide, followed by successive treatment, as described, with trichloroacetic acid and acetone, gives renin in good yield of a purity suitable for physiological studies and a good starting material for further purification. It contains 15 dog units per mg. N. 2. Further successive purification of this material with ethyl alcohol and ammonium sulfate gives a preparation containing 130 dog units per mg. N. The purest preparation hitherto reported (15) contained 16.0 to 20.8 units per mg. N. Preliminary Tiselius electrophoresis studies suggest homogeneity, but further studies to establish purity are in progress. 3. The properties of the most purified renin indicate that it is a protein. Its chemical and physiological properties correspond to those of the material in crude renal extract which induces an elevation of blood pressure when it is injected intravenously.

Calcium-45 binding by human prealbumin

1959 ◽  
Vol 14 (5) ◽  
pp. 859-860 ◽  
Author(s):  
Philip C. Johnson ◽  
William O Smith ◽  
Beulah Wulff
Keyword(s):  
Electrophoretic Mobility ◽  
Calcium Binding ◽  
Protein Separation ◽  
Gas Flow ◽  
Specific Activity ◽  
Tap Water ◽  
Paper Electrophoresis ◽  
Human Albumin ◽  
Binding Ability ◽  
Human Prealbumin

Calcium-45 binding by human albumin and prealbumin has been studied. Protein separation was performed by continuous paper electrophoresis. Calcium-45 was added to the fractions and the mixture incubated at 37℃ for 24 hours. Unbound calcium-45 was removed by dialysis against tap water for 16 hours, the pH of the fractions remained between 8.6–9.1 at all times. Aliquots of the fractions were then counted with a micromil gas flow counter. The greatest specific activity was found in several tubes preceding the albumin, i.e. with faster electrophoretic mobility than classical albumin. The calcium-binding ability of this fraction, which has been previously called ‘prealbumin,’ exceeded that of classical albumin by sixfold. Submitted on December 1, 1958

Preparation and Reactions of Trichloromethyl-Substituted Pyridine and Pyrimidine Derivatives

Synthesis ◽  
2018 ◽  
Vol 50 (20) ◽  
pp. 4071-4080 ◽  
Author(s):  
Reinhold Zimmer ◽  
Hans-Ulrich Reissig ◽  
Lina Unger ◽  
Matteo Accorsi ◽  
Christian Eidamshaus ◽  
...  
Keyword(s):  
Acetic Anhydride ◽  
Good Yield ◽  
Trichloroacetic Acid ◽  
Pyridine Derivative ◽  
Pyrimidine Derivatives ◽  
Methyl Group ◽  
Alternative Approach ◽  
Three Component Reaction ◽  
Trichloromethyl Group ◽  
Pyridine And Pyrimidine Derivatives

The three-component reaction of lithiated methoxyallene, nitriles and trichloroacetic acid gave three model β-keto enamides that were starting materials for the synthesis of trichloromethyl-substituted pyridine and pyrimidine N-oxide derivatives. Upon treatment with acetic anhydride, the methyl group of the prepared pyrimidine N-oxides was converted into an acetoxymethyl group by a Boekelheide rearrangement. A few typical experiments also revealed that the trichloromethyl group of the prepared pyrimidine N-oxides can be replaced by an alkoxy or a hydroxy group, or transformed into an arylthiomethyl group. An alternative approach to β-keto enamides via the corresponding β-keto enamines was also examined and provided the expected 4-hydroxy-6-(trichloromethyl)pyridine derivative in good yield.

Bindungsfähigkeit zytoplasmatischer Proteine. I. Farbstoffbindung

1958 ◽  
Vol 13 (8) ◽  
pp. 532-538 ◽  
Author(s):  
Ekkehard Kallee ◽  
Willmar Oppermann
Keyword(s):  
Adsorption Isotherm ◽  
Serum Proteins ◽  
Blood Stream ◽  
Bromophenol Blue ◽  
Cellular Membranes ◽  
Binding Ability ◽  
Cytoplasmic Proteins ◽  
Dye Binding ◽  
Rat Livers ◽  
Dialysis Technique

1. The dye-bindung capacity of cytoplasmic proteins obtained from mitochondria, microsomes and fluid cytoplasm of rat livers was estimated in dialysis experiments.As compared with serum proteins the binding of bromophenol blue to cytoplasmic proteins is in the same range as its affinity to serum proteins. The binding of these proteins corresponds to an adsorption isotherm.2. An application of a new dialysis technique — distribution dialysis — is described.3. The meanings of both the cellular membranes and dye-binding ability of cytoplasmic proteins for the uptake of substances from the blood stream into the interior of the cells are discussed.

A comparison of the Bradford and Lowry methods for the analysis of protein in chlorophyllous tissue

1982 ◽  
Vol 60 (7) ◽  
pp. 1046-1049 ◽  
Author(s):  
J. M. O. Eze ◽  
E. B. Dumbroff
Keyword(s):  
Trichloroacetic Acid ◽  
Leaf Tissue ◽  
Acid Precipitation ◽  
High Protein ◽  
Protein Yield ◽  
Large Protein ◽  
Leaf Proteins ◽  
Dye Binding ◽  
Bradford Assay ◽  
Significant Interference

A comparison of the dye-binding method of Bradford (1976) with the Folin phenol method of Lowry et al. (1951) for the assay of protein in chlorophyllous tissue is described. Additions of chlorophyll to protein standards precluded reliable determinations with either method. Absorbance readings were increased by nearly 20% with the Bradford assay and by over 400% using the Lowry procedure. Trichloroacetic acid precipitation of the proteins effectively eliminated any significant interference by chlorophyll; however, the practice of washing the protein precipitate with 80% acetone to remove residual chlorophyll resulted in large protein losses. Eluting leaf tissue with acetone prior to extraction of protein increased protein yield significantly. Both assay procedures provide satisfactory measurement of leaf proteins once chlorophyll is removed. The dye-binding method is fast, simple, and more sensitive, but the Lowry procedure gives better linearity at high protein concentrations and better stability once color has developed.

scholarly journals Total Protein Determination in Urine: Elimination of a Differential Response between the Coomassie Blue and Pyrogallol Red Protein Dye-binding Assays

2000 ◽  
Vol 46 (3) ◽  
pp. 392-398 ◽  
Author(s):  
Thomas Marshall ◽  
Katherine M Williams
Keyword(s):  
Total Protein ◽  
Control Level ◽  
Human Albumin ◽  
Differential Response ◽  
Urinary Protein ◽  
Total Protein Content ◽  
Binding Assays ◽  
Protein Determination ◽  
Pyrogallol Red ◽  
Dye Binding

Abstract Background: The total protein content of urine is a good index of renal function, but its determination is unreliable. Protein dye-binding assays are simple, but they characteristically lack a uniform response to different proteins. Methods: We investigated a differential response of the Sigma Microprotein Coomassie Brilliant Blue (CBB) and Pyrogallol Red-molybdate (PRM) protein dye-binding assays to urine, using human albumin, albumin/globulin, or urinary protein as calibrator. Results: The urine protein values (n = 60) obtained with the CBB assay were 110–13 500 mg/L (mean, 2390 mg/L) compared with 160–18 300 mg/L (mean, 3470 mg/L) obtained with the PRM assay (CBB:PRM protein concentration ratio, 0.46–0.88, mean, 0.69 ± 0.10). The differential response was highly reproducible as indicated by Sigma urine control Level 1 (within-day CBB:PRM ratio, 0.68 ± 0.02; between-day CBB:PRM ratio, 0.67 ± 0.04) and Sigma urine control Level 2 (within-day CBB:PRM ratio, 0.60 ± 0.01; between-day CBB:PRM ratio, 0.59 ± 0.02). The use of urinary protein as a calibrator (rather than human albumin) greatly improved the agreement between the assays when applied to urine (yCBB = 0.972xPRM − 16 vs yCBB = 0.685xPRM + 17). In studies using urine controls, this calibrator also improved agreement between the CBB, PRM, trichloroacetic acid (TCA), and benzethonium chloride protein methods and, to a lesser extent, agreement with the TCA-Ponceau S method. Conclusion: The use of a urinary protein calibrator improves the agreement between different methods used to determine total protein in urine.

Asialo von Willebrand Factor Enhances Platelet Adhesion to Vessel Subendothelium

1988 ◽  
Vol 60 (01) ◽  
pp. 030-034 ◽  
Author(s):  
Eva Bastida ◽  
Juan Monteagudo ◽  
Antonio Ordinas ◽  
Luigi De Marco ◽  
Ricardo Castillo
Keyword(s):  
Von Willebrand Factor ◽  
Platelet Adhesion ◽  
Human Albumin ◽  
Membrane Receptors ◽  
Shear Rates ◽  
High Shear Rates ◽  
Platelet Deposition ◽  
Platelet Spreading ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryNative von Willebrand factor (N-vWF) binds to platelets activated by thrombin, ADP or ristocetin. Asialo vWF (As-vWF) induces platelet aggregation in absence of platelet activators. N-vWF mediates platelet adhesion to vessel subendothelium at high shear rates. We have investigated the role of As-vWF in supporting platelet deposition to rabbit vessel subendothelium at a shear rate of 2,000 sec-1, using the Baumgartner perfusion system. We have studied the effects of the addition of As-vWF (from 2 to 12 μg/ml) to perfusates consisting of washed red blood cells, 4% human albumin and washed platelets. Our results show a significant increase in platelet deposition on subendothelium (p <0.01) in perfusions to which As-vWF had been added. Blockage of the platelet glycoproteins Ib and IIb/IIIa (GPIb and GPIIb/IIIa) by specific monoclonal antibodies (LJIb1 and LJCP8, respectively) resulted in a decrease of platelet deposition in both types of perfusates prepared with N-vWF and As-vWF. Our results indicate that As-vWF enhances platelet deposition to vessel subendothelium under flow conditions. Furthermore, they suggest that this effect is mediated by the binding of As-vWF to platelet membrane receptors, which in turn, promote platelet spreading and adhesion to the subendothelium.

Treatment of Hemophilia A with a Highly Purified Factor VIII Concentrate Prepared by Anti-FVIIIc Immunoaffinity Chromatography

1992 ◽  
Vol 67 (01) ◽  
pp. 019-027 ◽  
Author(s):  
Joseph E Addiego ◽  
Edward Gomperts ◽  
Liu Shu-Len ◽  
Patricia Bailey ◽  
Suzanne G Courter ◽  
...  
Keyword(s):  
Factor Viii ◽  
Hemophilia A ◽  
Virus Transmission ◽  
Human Albumin ◽  
Immunoaffinity Chromatography ◽  
Exchange Chromatography ◽  
Pharmacokinetic Studies ◽  
Enveloped Viruses ◽  
Clinically Significant ◽  
Detergent Mixture

SummaryTo reduce the risk of pathogenic virus transmission associated with the therapeutic administration of plasma-derived antihemophilic factor (FVIIIc), a process utilizing anti-FVIIIc immunoaffinity chromatography to isolate FVIIIc has been developed. In addition, the starting cryoprecipitate solution has been treated with an organic solvent/detergent mixture to inactivate lipid-enveloped viruses. A final ion exchange chromatography step is used to further remove contaminants, e.g., anti-FVIIIc antibody, potentially leached with FVIIIc during the immunoaffinity step. The purified FVTII is stabilized for lyophili-zation and storage by the addition of human albumin. The monoclonal anti-FVIIIc antibody used in the immunoaffinity step of the process is not detectable in the final preparation. Viral reduction studies performed at specific steps of the process demonstrate that 11 logs of human immunodeficiency virus (HIV) and greater than 4-5 logs of other lipid-enveloped viruses are inactivated within the first 30 s of exposure to the solvent/ detergent mixture and 4-5 logs of various model viruses, e. g. Endomyocarditis virus (EMC), are physically removed during washing of the immunoaffinity column. The lyophilized product is reconstituted using sterile water in a matter of seconds.The pharmacokinetics of Hemofil® M were compared to those obtained using a standard heat-treated concentrate (Hemofil® CT) in five severe factor VIII deficient hemophiliacs in a randomized, cross-over study. No statistically significant differences were observed in mean half life (p >0.6) or median recovery (p = 0.4) between the two preparations. No clinically significant adverse effects were observed in patients receiving either FVIII preparation.In addition, 43 patients at 18 different centers underwent pharmacokinetic studies, with a nominal dose of 50 u/kg FVIIIc Hemofil® M. The mean recovery was 103.6%, and the t 1/2 was 14.6 h. The recovery of FVIII in this group was as expected, providing an increase of assayed FVIII of approximately 2% per unit of FVTII/kg infused.Clinical trials using Hemofil® M have been initiated in 124 hemophilia A patients. The safety and efficacy of Hemofil® M has been established. To date, 0 of 60 patients tested have seroconverted to HIV. None of the previously untreated patients show clinical or laboratory evidence of Non-A, Non-B hepatitis (NANB), with 21 patients remaining negative as far as presence of antibodies to the Hepatitis C virus (a-HCV negative) at least 6 months after the initial infusion. There is no evidence of neoantigenicity, evidenced by seroconversion to murine antibody. An 8.7% (2 of 23) prevalence of anti-FVIIIc inhibitor development has been observed in previously untreated patients with FVIIIc⩽3%, receiving only the monoclonally purified solvent/ detergent treated FVIII concentrate while on study and on poststudy surveillance. All patients demonstrated clinical hemostasis following product use for either on demand bleeding or surgical prophylaxis.

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