scholarly journals Fast Surface Immobilization of Native Proteins through Metal-Free Amino-yne Click Bioconjugation

2019 ◽  
Author(s):  
Yiru Zhang ◽  
Jianlei Shen ◽  
Rong Hu ◽  
Shi Xiujuan ◽  
Xianglong Hu ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Free Amino ◽  
Surface Immobilization ◽  
Ethynyl Group ◽  
Metal Free ◽  
Native Proteins ◽  
Functionalized Surfaces ◽  
Bulk Solutions

In this work, we use a spontaneous and catalyst-free amino-yne click bioconjugation to generate activated ethynyl group functionalized surfaces for fast immobilizing native proteins and cells. Biomolecules, such as bovine serum albumin (BSA), human IgG and peptide of C(RGDfK), could be covalently immobilized on the surfaces in as short as 30 min. Notably, the bioactivity of the anchored biomolecules remain intact, which is verified by efficiently capturing target antibodies and cells from the bulk solutions. This strategy represents an alternative for highly efficient surface biofunctionalization.

scholarly journals Fast Surface Immobilization of Native Proteins through Metal-Free Amino-yne Click Bioconjugation

2019 ◽  
Author(s):  
Yiru Zhang ◽  
Jianlei Shen ◽  
Rong Hu ◽  
Shi Xiujuan ◽  
Xianglong Hu ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Free Amino ◽  
Surface Immobilization ◽  
Ethynyl Group ◽  
Metal Free ◽  
Native Proteins ◽  
Functionalized Surfaces ◽  
Bulk Solutions

In this work, we use a spontaneous and catalyst-free amino-yne click bioconjugation to generate activated ethynyl group functionalized surfaces for fast immobilizing native proteins and cells. Biomolecules, such as bovine serum albumin (BSA), human IgG and peptide of C(RGDfK), could be covalently immobilized on the surfaces in as short as 30 min. Notably, the bioactivity of the anchored biomolecules remain intact, which is verified by efficiently capturing target antibodies and cells from the bulk solutions. This strategy represents an alternative for highly efficient surface biofunctionalization.

SuFEx: a metal-free click ligation for multivalent biomolecules

2017 ◽  
Vol 15 (7) ◽  
pp. 1549-1553 ◽  
Author(s):  
Alessandro Dondoni ◽  
Alberto Marra
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Metal Free ◽  
Sulfonyl Fluoride

Sulfonamide-linked glycoclusters and sulfamate-linked BSA-PEG were prepared by coupling a sugar sulfonyl fluoride with a calixarene tetra-amine and a PEG-fluorosulfate with native bovine serum albumin, respectively.

Effect of a Test Meal, without and with Protein, on Muscle and Plasma Free Amino Acids

1990 ◽  
Vol 79 (4) ◽  
pp. 331-337 ◽  
Author(s):  
J. Bergström ◽  
P. Fürst ◽  
E. Vinnars
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Free Amino Acids ◽  
Free Amino ◽  
Essential Amino Acids ◽  
Muscle Concentration ◽  
Protein Rich

1. The effect of a protein-free meal and a protein-rich meal on the concentration of free amino acids in plasma and muscle tissue was studied in eight healthy subjects. The energy content of the protein-free meal was 3800 kJ. The protein-rich meal was identical in composition except that 50 g of bovine serum albumin was added. Plasma and samples from the quadriceps femoris muscle (percutaneous muscle biopsy) for amino acid determination were collection before and at 1, 3, 5 and 7 h after the meal. 2. After the protein-free meal the concentrations of most essential amino acids and of some non-essential amino acids in plasma decreased continuously below basal levels at 5–7 h. The muscle concentration of essential amino acids fell too, reaching its nadir 3–5 h after the meal. The decrease in plasma amino acid concentration was smaller than the decrease in muscle concentration for all essential amino acids except phenylalanine. 3. The concentrations of most amino acids in plasma increased transiently 1 and 3 h after the protein-rich meal; histidine and several non-essential amino acids fell below the basal levels at 5–7 h after the meal. In muscle, threonine, valine, leucine, lysine and alanine were increased at 1 and 3 h after the protein-rich meal; isoleucine, serine and glycine fell below the basal level after 5 and 7 h. For the essential amino acids except threonine and lysine, the increase in plasma concentration was greater than the increase in muscle concentration. 4. Correlations were observed between the relative content of the essential amino acids in the bovine serum albumin and the increase in concentration of these amino acids in plasma and muscle. Methionine and isoleucine, which had the lowest content in bovine serum albumin, fell below basal levels in plasma and (for isoleucine) in muscle 5–7 h after the meal, suggesting that these two amino acids might have been limiting for protein synthesis. 5. Amino acid analysis in plasma and muscle samples taken postprandially should be evaluated with caution considering the strong influence of meal composition on plasma and muscle free amino acid profiles.

scholarly journals Anti-flavin antibodies

1987 ◽  
Vol 242 (1) ◽  
pp. 89-95 ◽  
Author(s):  
M J Barber ◽  
D C Eichler ◽  
L P Solomonson ◽  
B A Ackrell
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Succinate Dehydrogenase ◽  
Bovine Serum ◽  
Deae Cellulose ◽  
Double Diffusion ◽  
Cross Reactivity ◽  
Rabbit Serum ◽  
Native Proteins ◽  
Increased Sensitivity

Antibodies were elicited to FAD by using the hapten N-6-(6-aminohexyl)-FAD conjugated to the immunogenic carrier protein bovine serum albumin. Cross-reactivity was determined by Ouchterlony double-diffusion analysis with N-6-(6-aminohexyl)-FAD coupled to rabbit serum albumin. Anti-FAD IgG was partially purified by (NH4)2SO4 precipitation followed by DEAE-cellulose/CM-cellulose and bovine serum albumin-agarose chromatography. The partially purified anti-FAD IgG fraction failed to inhibit the catalytic activities of the flavin-containing enzymes nitrate reductase, xanthine oxidase and succinate dehydrogenase, whereas enzyme activity could be inhibited by addition of antibodies elicited against the native proteins. However, the partially purified anti-FAD IgG fraction could be used as a highly sensitive and specific probe to detect proteins containing only covalently bound flavin, such as succinate dehydrogenase, p-cresol methylhydroxylase and monoamine oxidase, by immuno-blotting techniques. Detection limits were estimated to be of the order of femtomolar concentrations of FAD with increased sensitivity for the 8 alpha-N(3)-histidyl linkage compared with 8 alpha-O-tyrosyl substitution.

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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