Choriogonadotropin measured with the Tandem-E immunoenzymetric assay system.

1985 ◽  
Vol 31 (3) ◽  
pp. 441-444 ◽  
Author(s):  
J L Bock ◽  
J Furgiuele ◽  
J C Segen
Keyword(s):  
Alkaline Phosphatase ◽  
Linear Response ◽  
Beta Subunit ◽  
Good Precision ◽  
False Positive Result ◽  
Range Correlation ◽  
Sandwich Type ◽  
Plastic Bead ◽  
Substrate Hydrolysis ◽  
Higher Sensitivity

Abstract We evaluated the Hybritech Tandem-E procedure for quantifying choriogonadotropin (hCG) in human serum. In this "sandwich"-type assay, two monoclonal antibodies directed against different regions of the hCG molecule are used, one coated on a plastic bead, the second conjugated to alkaline phosphatase. The assay can detect as little as 1.0 int. unit of the hormone per liter, shows a linear response up to at least 200 int. units/L, and has good precision. By prolonging the incubations for formation of the sandwich and for substrate hydrolysis, one can achieve higher sensitivity at the expense of a narrower linear range. Correlation with a conventional radioimmunoassay for the beta subunit of hCG was generally excellent, but in one instance the Tandem-E gave an apparently false positive result.

A Novel Electrochemical Immunosensor Based on DNA/Fe3O4 (Core) /ZrO2（Shell）one Dimension Magnetic Probes for Carcino-Embryonic Antigen

2011 ◽  
Vol 80-81 ◽  
pp. 200-206 ◽  
Author(s):  
Fu Tao Hu ◽  
Yuan Zao Wu ◽  
Yu Ting Cao ◽  
Ning Gan
Keyword(s):  
Electrochemical Immunosensor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Good Precision ◽  
Current Signal ◽  
Separation And Purification ◽  
Rapid Separation ◽  
Catalytic Current ◽  
Nano Gold ◽  
Modified Dna ◽  
Sandwich Type

A novel and sensitive electrochemical enzyme-linked immunosorbent assay (ELISA) system of Hydroquinones (HQ)- peroxide hydantoin (CP) -horseradish-peroxidase (HRP) by using magnetic DNA linked core-shell Fe3O4@ZrO2（ZMPs）nanoparticles as labels and signal amplification was established. In the presence of carcinoembryonic antigen (CEA) analyte, the sandwich-type immunocomplex could be formed between HRP labeled CEA second antibody（CEA Ab2） modified DNA-ZMPs probes (DNA / (ZMPs-HRP-CEA Ab2) n) and nano-gold CEA modified glassy carbon electrode (GCE | CEA Ab1) as an immune electrode.The concentration of CEA was determined based on the current signal, which was generated in the reaction between HQ and CP catalyzed by HRP on the sandwich-type immunocomplex. ZMPs nanoparticles as label material can not only perform the rapid separation and purification of signal antibody on magnetic field, but also immensely enhance the labeled capacity of HRP-anti-CEA which can amplify the catalytic current signal. Furthermore, the electrode modification process was not required. The method provided a linear response range between 0.01 and 200 ng·mL-1with a detection limit of 4 pg·mL-1.Moreover, the proposed electrochemical ELISA method exhibited good precision, high sensitivity and acceptable reproducibility, and could be further developed for clinical detections of CEA and other biomarkers.

AutoAnalyzer Assay for Serum Alkaline Phosphatase Activity, with Sodium Thymolphthalein Monophosphate as Substrate

1973 ◽  
Vol 19 (1) ◽  
pp. 103-105 ◽  
Author(s):  
Gary J Proksch ◽  
Dean P Bonderman ◽  
John A Griep
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Serum Alkaline Phosphatase ◽  
Flow Diagram ◽  
Good Precision ◽  
Serum Alkaline Phosphatase Activity ◽  
Manual Method ◽  
Automated Method ◽  
Simple Flow

Abstract An automated method is described for determining alkaline phosphatase activity in serum, with use of sodium thymolphthalein monophosphate as the substrate. The system makes use of standard AutoAnalyzer components, has a simple flow diagram, and does not involve dialysis. The method has good precision, and the results correlate well with those from the manual method of which it is an adaptation

Phosphatase Activity in Homogenates from Normal and Poliovirus Infected Tissue Cultures

1963 ◽  
Vol 18 (11) ◽  
pp. 903-912 ◽  
Author(s):  
E. Frank Deig ◽  
Louis P. Gebhardt
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Substrate Concentration ◽  
Initial Substrate ◽  
Cell Homogenate ◽  
Normal Monkey ◽  
Proportional Increase ◽  
Substrate Hydrolysis ◽  
Assay Conditions

A study was made on the total ATP-ase, alkaline phosphatase, and glucose-6-phosphatase activity in homogenates prepared from normal monkey kidney cells in tissue culture. A characterization of such activity under standardized assay conditions revealed that in comparable amounts of cell homogenate ATP-ase was 8—10 times as active as alkaline phosphatase and glucose-6-phosphatase. Substrate hydrolysis was brought about by the activity of each enzyme at a rate which was constant with respect to time. The amount of substrate hydrolyzed, in each case, was directly proportional to the concentration of homogenate used in the assay system. Within certain limits this was also true with respect to substrate concentration. However, with increasing amounts of substrate and a constant amount of cell homogenate a point was reached where the activity of each enzyme became independent of initial substrate concentration. Under one set of assay conditions the total ATP-ase in cell homogenates was shown to be optimally active at approximately pH 7 while under another the enzyme showed a nearly directly proportional increase in activity per unit time between pH 4 and pH 10. Using standardized assay conditions such ATP-ase activity was markedly enhanced over that found in similar systems lacking added calcium and magnesium ion. No further stimulation of the activity of this enzyme, or of alkaline phosphatase and glucose-6-phosphatase, resulted from their interaction with 2.4-dinitrophenol. The substrate hydrolysis produced by the activity of each enzyme in cell homogenate preparations resulted from irreversible reactions. Such homogenates were incapable of carrying out an exchange reaction of radioactive orthophosphate with the phosphorous from any of the substrates used in this study.

BIOASSAY OF PARATHYROID HORMONE IN RATS BY DETERMINATION OF PLASMA CALCIUM, URINARY 32P EXCRETION AND SERUM ALKALINE PHOSPHATASE

1966 ◽  
Vol 35 (3) ◽  
pp. 229-238 ◽  
Author(s):  
R. J. TREACHER
Keyword(s):  
Alkaline Phosphatase ◽  
Parathyroid Hormone ◽  
Serum Calcium ◽  
Calcium Concentration ◽  
Serum Alkaline Phosphatase ◽  
Plasma Calcium ◽  
Significant Alteration ◽  
Good Precision ◽  
Serum Calcium Concentration

SUMMARY Methods for assay of parathyroid hormone based on an increase in serum calcium concentration, urinary 32P excretion and serum alkaline phosphatase elevation in parathyroidectomized rats have been compared and modifications introduced to improve sensitivity, precision, speed and ease of manipulation. Both the serum calcium and urinary 32P assay gave good precision (mean λ = 0·23 and 0·29, respectively) but by the serum calcium method less than 10 USP units of parathyroid hormone could not be detected, whereas the phosphaturic assay detects as little as 0·5 USP unit. Both assays are simple to perform and each requires only 2 days to complete. They can be combined in a single design using the same animals. Assays based on serum alkaline phosphatase levels in parathyroidectomized rats were not successful since it was impossible to produce a significant alteration in serum alkaline phosphatase by the administration of parathyroid hormone.

scholarly journals Response in nutritionally related blood metabolites, carcass traits and primal pork cuts of slow growing Windsnyer pigs fed on varying levels of potato hash silage

1970 ◽  
Vol 48 (4) ◽  
pp. 770-776
Author(s):  
C.N. Ncobela ◽  
A.T. Kanengoni ◽  
M. Chimonyo
Keyword(s):  
Alkaline Phosphatase ◽  
Linear Relationship ◽  
Total Protein ◽  
Linear Response ◽  
Carcass Traits ◽  
Carcass Weight ◽  
Muscle Area ◽  
Eye Muscle ◽  
Inclusion Level ◽  
Quadratic Relationship

The response of Windsnyer pigs to diets containing varying levels of potato hash silage in nutritionally related blood biochemistry, carcass traits and primal pork was estimated. Thirty-six growing clinically healthy male Windsnyer pigs with an initial weight of 36 kg ± 4.89 (mean ± standard deviation (SD)) were randomly assigned to six experimental diets containing 0, 80, 160, 240, 320, and 400 potato hash silage g/kg dry matter (DM). Experimental diets were derived from mixing a summit diet containing no potato hash silage and a dilution diet containing 400 g potato hash silage/kg in various proportions. Pigs were allowed ad libitum access to diets and water. There was no relationship between inclusion levels of potato hash silage and albumin: globulin ratio, total protein, and uric acid. As inclusion levels of potato hash silage varied, there was a positive linear relationship between silage and albumin concentration. Globulin concentration had a positive quadratic relationship with the inclusion of potato hash silage. Inclusion levels of potato hash silage resulted in a positive quadratic relationship in alkaline phosphatase. There was a negative linear response in warm carcass weight and cold carcass weight to inclusion levels of silage. A negative linear response was observed in dressing percentage. Different inclusion levels of potato hash silage caused a positive quadratic relationship in cooler shrink. There were negative linear relationships between inclusion of potato hash silage with shoulder fat, carcass length and backfat thickness. There was a negative linear relationship between eye muscle area and inclusion level of ensiled potato hash. There was a positive quadratic relationship between hindquarter length (HQL) and inclusion levels of silage. The observed linear relationship between hindquarter circumference (HQC) and inclusion levels of potato hash silage was negative. There is a need to predict the optimum inclusion level of potato hash silage without compromising the healthiness and carcass yield of pigs.Keywords: Alkaline phosphatase, backfat thickness, cold carcass weight, cooler shrink, dorsal fat thickness, eye muscle area, hindquarter circumference, total protein

Amperometric Biosensors Using Different Alcohol Oxidases

2019 ◽  
Vol 891 ◽  
pp. 90-95
Author(s):  
Manatsapon Tipmanee ◽  
Saipin Thanachasai
Keyword(s):  
Detection Limit ◽  
Pichia Pastoris ◽  
Linear Response ◽  
Current Response ◽  
Initial Value ◽  
Amperometric Biosensors ◽  
Different Sources ◽  
Higher Sensitivity

Amperometric biosensors were fabricated by immobilizing alcohol oxidases (AOX) from two different sources onto glutaraldehyde (GA)-activated supports. Alcohol oxidases fromHansenulasp. and fromPichia pastoriswere employed for immobilization. The biosensor with AOX fromHansenulasp. showed a linear response to ethanol in the concentration range of 0.1-0.6 mM with a sensitivity of 88.534 µA mM-1cm-2and a detection limit of 0.1 mM (S/N=3). In comparison, the biosensor with AOX fromP. pastorisshowed a linear response from 0.1-0.5 mM ethanol with a sensitivity of 76.886 µA mM-1cm-2and a detection limit of 0.1 mM. The study of stability of biosensors revealed that after 90 measurements, the biosensor with AOX fromHansenulasp. retained 97% of its original current response whereas the current response of the biosensor with AOX fromP. pastorisdecreased to 81% of its initial value. The biosensor with AOX fromHansenulasp. demonstrated slightly higher sensitivity and stability than the biosensor with AOX fromP. pastoris.

scholarly journals Studies on alkaline phosphatase. Phosphorylation of calf intestinal alkaline phosphatase by 32P-labelled pyrophosphate

1968 ◽  
Vol 107 (2) ◽  
pp. 279-283 ◽  
Author(s):  
H N Fernley ◽  
Sylvia Bisaz
Keyword(s):  
Alkaline Phosphatase ◽  
Intestinal Mucosa ◽  
Active Sites ◽  
Stopped Flow ◽  
Serine Residue ◽  
Intestinal Alkaline Phosphatase ◽  
Calf Intestinal Alkaline Phosphatase ◽  
Substrate Hydrolysis

1. A purified preparation of alkaline phosphatase from calf-intestinal mucosa was phosphorylated by 32P-labelled PPi at a serine residue on the enzyme. Under the conditions employed, up to 0·15μm-labelled sites were obtained from 1μm-[32P]PPi. 2. The phosphorylated enzyme was labile, the rate of dephosphorylation being similar to the overall rate of substrate hydrolysis. 3. A stopped-flow technique was used to determine the number of phosphomonoesterase active sites, which agreed with the number of 32P-labelled sites. 4. It is concluded that calf-intestinal alkaline phosphatase is both a phosphomonoesterase and a pyrophosphatase.

scholarly journals Effect of Long-Term Hypodynamy on Alkaline Phosphatase Activity of Small Intestine in Japanese Quail Chicks

2007 ◽  
Vol 76 (3) ◽  
pp. 333-338
Author(s):  
Ľ. Lenhardt ◽  
V. Cigánková ◽  
V. Almášiová ◽  
K. Holovská ◽  
P. Škrobánek ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Small Intestine ◽  
Japanese Quail ◽  
Simulated Microgravity ◽  
Intestinal Function ◽  
Simulated Weightlessness ◽  
Experimental Animals ◽  
Functional Development ◽  
Higher Sensitivity

The functional development of the small intestine was investigated in Japanese quail chicks subjected to simulated microgravity (hypodynamy) on the second day after hatching and reared under these conditions to 63 days of age. On days 5, 7, 14, 21, 28, 35, 42, 56 and 63 the activity of brush-border-bound alkaline phosphatase (AP) in the duodenum and jejunum were determined in experimental animals as well as in control quail chicks housed in a floor box during these periods. As compared with control quails the experimental animals displayed a significantly increased enzyme activity until day 42 in the duodenum and day 35 in the jejunum (P < 0.001) whereas in older quails no significant enzymatic differences between these groups was found. However, a decrease in food consumption due to a partial physical constraint cannot be excluded. Moreover, the results suggested that the activity of AP in the control birds did not change substantially during all the periods examined. In contrast, in older hypodynamy quail the AP activity significantly decreased in the duodenum on days 56 and 63 and in the jejunum on days 42, 56 and 63, respectively. These results indicate that a) the enhanced intestinal function in early periods of life may reflect the higher sensitivity of small intestine to simulated weightlessness, b) the decrease of the AP activity in older animals to the level of controls might be considered as a part of intestinal mechanisms involved in adaptation of quail chicks to long-term hypodynamy, c) different activity of AP in the small intestine of Japanese quail may not have resulted only from hypodynamy but also due to decreased food intake.

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