Interlaboratory Study of the IFCC Method for Alanine Aminotransferase Performed with Use of a Partly Purified Reference Material

1992 ◽  
Vol 38 (12) ◽  
pp. 2365-2371 ◽  
Author(s):  
F Schiele ◽  
J Muller ◽  
E Colinet ◽  
G Siest ◽  
P Arzoglou ◽  
...  
Keyword(s):  
Reference Material ◽  
Bovine Serum Albumin ◽  
Confidence Interval ◽  
Serum Albumin ◽  
Enzyme Activities ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Bovine Serum ◽  
Specific Activity ◽  
Interlaboratory Trial

Abstract We present the results of a study on performance of a reference material for alanine aminotransferase (ALT, EC 2.6.1.2) and the corresponding IFCC-approved method in an interlaboratory trial involving 13 laboratories. The ALT material was partly purified from pig heart (specific activity, 150 kU/g) and was essentially free of six potentially contaminating enzyme activities, including aspartate aminotransferase (EC 2.6.1.1). The partly purified ALT was lyophilized in a triethanolamine-buffered matrix, pH 6.4, containing bovine serum albumin and saccharose. Under these conditions, the predicted yearly loss of activity was 0.02% at 4 degrees C and < 0.01% at -20 degrees C. The final blank-corrected results of the accepted set of data gave a mean (SD) of 128.5 (5.1) U/L. The among-laboratory SD was 4.6 U/L and the within-laboratory SD was 2.0 U/L. The certified ALT catalytic concentration in the reconstituted material was 129 U/L with a 0.95 confidence interval of +/- 4 U/L.

Purification of Progesterone Antiserum for Radioimmunoassay, by Chromatography on QAE-Sephadex A-50

1973 ◽  
Vol 19 (9) ◽  
pp. 1027-1030 ◽  
Author(s):  
Frank H Bodley ◽  
Alcide Chapdelaine ◽  
Kenneth D Roberts
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Column Chromatography ◽  
Bovine Serum ◽  
Specific Activity ◽  
Antibody Preparation ◽  
Covalent Coupling

Abstract Antibodies were generated in sheep after administration of the antigen, progesterone-11α-succinyl-bovine serum albumin. This antiserum was purified 10-fold by treatment with "Aerosil" and column chromatography on "QAE-Sephadex A-50." In addition to an increased specific activity (cpm progesterone bound/g of protein) the purified antibody fraction was less cross-reactive with pregnenolone and desoxycorticosterone than was the crude antiserum. A sensitivity of 10 pg of progesterone was obtained with the purified antibody preparation, compared to 25 pg for the crude antiserum. This method of purification of antisera was devised in order to concentrate the antibody fraction before covalent coupling to insoluble supports such as activated arylamine-glass particles.

scholarly journals Enzyme Stability in Nanoparticle Preparations Part 1: Bovine Serum Albumin Improves Enzyme Function

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4593
Author(s):  
Jason Thomas Duskey ◽  
Federica da Ros ◽  
Ilaria Ottonelli ◽  
Barbara Zambelli ◽  
Maria Angela Vandelli ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Specific Activity ◽  
Enzyme Stability ◽  
Polymeric Nanoparticles ◽  
Release Kinetics ◽  
Titration Calorimetry ◽  
Therapeutic Effects

Enzymes have gained attention for their role in numerous disease states, calling for research for their efficient delivery. Loading enzymes into polymeric nanoparticles to improve biodistribution, stability, and targeting in vivo has led the field with promising results, but these enzymes still suffer from a degradation effect during the formulation process that leads to lower kinetics and specific activity leading to a loss of therapeutic potential. Stabilizers, such as bovine serum albumin (BSA), can be beneficial, but the knowledge and understanding of their interaction with enzymes are not fully elucidated. To this end, the interaction of BSA with a model enzyme B-Glu, part of the hydrolase class and linked to Gaucher disease, was analyzed. To quantify the natural interaction of beta-glucosidase (B-Glu,) and BSA in solution, isothermal titration calorimetry (ITC) analysis was performed. Afterwards, polymeric nanoparticles encapsulating these complexes were fully characterized, and the encapsulation efficiency, activity of the encapsulated enzyme, and release kinetics of the enzyme were compared. ITC results showed that a natural binding of 1:1 was seen between B-Glu and BSA. Complex concentrations did not affect nanoparticle characteristics which maintained a size between 250 and 350 nm, but increased loading capacity (from 6% to 30%), enzyme activity, and extended-release kinetics (from less than one day to six days) were observed for particles containing higher B-Glu:BSA ratios. These results highlight the importance of understanding enzyme:stabilizer interactions in various nanoparticle systems to improve not only enzyme activity but also biodistribution and release kinetics for improved therapeutic effects. These results will be critical to fully characterize and compare the effect of stabilizers, such as BSA with other, more relevant therapeutic enzymes for central nervous system (CNS) disease treatments.

Einbau von [32P]Orthophosphat in DNA der Mäuse-Milz unter Antigen-Einfluß / Incorporation of [32P]Orthophosphate into DNA of the Mouse Spleen under Influence of Antigen

1980 ◽  
Vol 35 (7-8) ◽  
pp. 599-604
Author(s):  
Wilhelm Uecker ◽  
Diethard Kubsch ◽  
Dieter Lutz ◽  
Hans Kröger
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Dna Synthesis ◽  
Satellite Dna ◽  
Bovine Serum ◽  
Specific Activity ◽  
Main Band ◽  
Relative Specific Activity ◽  
Specific Activities ◽  
Observation Period

Antigens cause an increase of the DNA synthesis in the spleen of mice as shown by the in­corporation of [32P]orthophosphate and [3H]thymidine. In the present paper, the incorporation of [32P]orthophosphate into the single deoxymononucleotides has been studied under influence of the antigen bovine serum albumin. For this purpose, the labelled DNA was decomposed to the deoxymononucleotides and their specific activities determined. Using [32P]orthophosphate, it was found that the activity of the DNA increased continuously during an observation period of 8 h. While after one hour the relative specific activity of dTMP was highest and that of dAMP lowest, the relative specific activities of the deoxymononucleotides had become equal after 8 h. Under the influence of bovine serum albumin, the incorporation of [32P]orthophosphate increased in the main band DNA as well as in the satellite DNA. The antigen had no effect on the distribution of the specific activities of the deoxymononucleotides.

Proposals for Standardization of Total Protein Assays

1968 ◽  
Vol 14 (12) ◽  
pp. 1147-1159 ◽  
Author(s):  
Theodore Peters
Keyword(s):  
Reference Material ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Total Protein ◽  
Bovine Serum ◽  
Dry Weight ◽  
Total Serum Protein ◽  
Total Serum ◽  
Reference Preparation ◽  
Protein Assays

Abstract The Standards Committee of the AACC presents a discussion of the nature of total serum protein and the problems associated with its determination. Proposals are made that protein be defined as polypeptide material, and that a reference preparation be promulgated for interlaboratory use. The reference material suggested is bovine serum albumin, produced to rigid specifications and distributed as a stable 7% (w/v) solution, the concentration of which has been established by careful dry weight assay. Comments of readers are invited.

Relative stabilities of purified human mitochondrial and cytoplasmic isoenzymes of aspartate aminotransferase in lyophilized materials.

1979 ◽  
Vol 25 (5) ◽  
pp. 659-664 ◽  
Author(s):  
E J Sampson ◽  
S S McKneally ◽  
V S Whitner ◽  
C A Burtis ◽  
D D Bayse
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Aspartate Aminotransferase ◽  
Bovine Serum ◽  
Degradation Process ◽  
Storage Conditions ◽  
Apparent Difference ◽  
Reaction Solution ◽  
The Matrix ◽  
The Stability

Abstract Eight different pools of purified human mitochondrial and cytoplasmic isoenzymes of aspartate aminotransferase were prepared, to examine the effects of the following matrix variables: the matrix support material (bovine serum albumin and polyvinylpyrrolidone), endogenous pyridoxal concentration, and azide as an antimicrobial preservation. Storage temperatures of 25 and 37 degrees C were used as a rapid and convenient means of accelerating the degradation process. Activity of the enzyme was measured with and without pyridoxal in the reaction solution. We found that the mitochondrial isoenzyme was consistently more labile than the cytoplasmic isoenzyme under identical storage conditions. Both isoenzymes were more stable in matrixes containing bovine serum albumin than in those containing polyvinylpyrrolidone. No apparent difference in the stability of either isoenzyme was observed at matrix pyridoxal concentrations of 15 micromol/L and 150 micromol/L. Only the mitochondrial isoenzyme in matrixes containing bovine serum albumin and 15 micromol of pyridoxal per liter had increased activity (about 9%) when pyridoxal was added to the enzymatic reagent. The amount of activity in reconstituted specimens did not apparently change after 72 h at 4 degrees C.

Development of bovine serum albumin certified reference material

2011 ◽  
Vol 400 (10) ◽  
pp. 3443-3449 ◽  
Author(s):  
Liqing Wu ◽  
Bin Yang ◽  
Jiaming Bi ◽  
Jing Wang
Keyword(s):  
Reference Material ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Certified Reference Material ◽  
Bovine Serum

Improvement of an artificial test substrate technique for evaluation of em immunocytochemical labeling

1987 ◽  
Vol 45 ◽  
pp. 762-763
Author(s):  
G. D. Gagne ◽  
M. F. Miller
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Artificial Substrate ◽  
Chemical Fixation ◽  
As If

We recently described an artificial substrate system which could be used to optimize labeling parameters in EM immunocytochemistry (ICC). The system utilizes blocks of glutaraldehyde polymerized bovine serum albumin (BSA) into which an antigen is incorporated by a soaking procedure. The resulting antigen impregnated blocks can then be fixed and embedded as if they are pieces of tissue and the effects of fixation, embedding and other parameters on the ability of incorporated antigen to be immunocyto-chemically labeled can then be assessed. In developing this system further, we discovered that the BSA substrate can also be dried and then sectioned for immunolabeling with or without prior chemical fixation and without exposing the antigen to embedding reagents. The effects of fixation and embedding protocols can thus be evaluated separately.

Stability of Prostacyclin in Human Plasma and Whole Blood: Studies on the Protective Effect of Albumin

1981 ◽  
Vol 46 (03) ◽  
pp. 645-647 ◽  
Author(s):  
M A Orchard ◽  
C Robinson
Keyword(s):  
Platelet Aggregation ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Protective Effect ◽  
Whole Blood ◽  
Bovine Serum ◽  
Fatty Acid Content ◽  
Krebs Solution ◽  
Antiaggregatory Activity ◽  
Free Plasma

SummaryThe biological half-life of prostacyclin in Krebs solution, human cell-free plasma or whole blood was measured by bracket assay on ADP-induced platelet aggregation. At 37°C, pH 7.4, plasma and blood reduced the rate of loss of antiaggregatory activity compared with Krebs solution. The protective effect of plasma was greater than that of whole blood. This effect could be partially mimicked by the addition of human or bovine serum albumin to the Krebs solution. The stabilisation afforded by human serum albumin was dependent on the fatty acid content of the albumin, although this was less important for bovine serum albumin.

RADIOIMMUNOASSAY OF OESTRONE AND OESTRADIOL IN THE PERIPHERAL PLASMA OF THE DOMESTIC FOWL IN VARIOUS PHYSIOLOGICAL STATES AND OF HYPOPHYSECTOMIZED AND OVARIECTOMIZED FOWLS

1974 ◽  
Vol 75 (1) ◽  
pp. 133-140 ◽  
Author(s):  
B. E. Senior
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Diethyl Ether ◽  
Bovine Serum ◽  
Domestic Fowl ◽  
Laying Hens ◽  
Peripheral Plasma ◽  
The Mean ◽  
Precision And Accuracy ◽  
Chicken Ovary

ABSTRACT A radioimmunoassay was developed to measure the levels of oestrone and oestradiol in 0.5–1.0 ml of domestic fowl peripheral plasma. The oestrogens were extracted with diethyl ether, chromatographed on columns of Sephadex LH-20 and assayed with an antiserum prepared against oestradiol-17β-succinyl-bovine serum albumin using a 17 h incubation at 4°C. The specificity, sensitivity, precision and accuracy of the assays were satisfactory. Oestrogen concentrations were determined in the plasma of birds in various reproductive states. In laying hens the ranges of oestrone and oestradiol were 12–190 pg/ml and 29–327 pg/ml respectively. Levels in immature birds, in adult cockerels and in an ovariectomized hen were barely detectable. The mean concentrations of oestrone and oestradiol in the plasma of four non-laying hens (55 pg/ml and 72 pg/ml respectively) and one partially ovariectomized hen (71 pg/ml and 134 pg/ml respectively) were well within the range for laying hens. It is evident that the large, yolk-filled follicles are not the only source of oestrogens in the chicken ovary.

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