Counterimmunoelectrophoresis in determination of prostatic acid phosphatase in human serum.

1978 ◽  
Vol 24 (1) ◽  
pp. 140-142 ◽  
Author(s):  
A G Foti ◽  
J F Cooper ◽  
H Herschman

Abstract We evaluated counterimmunoelectrophoresis for use in measuring prostatic acid phosphatase in detection of prostatic cancer. After staining for acid phosphatase, we could detect as little as 0.3 ng of purified enzyme standard complexed with antibody by this technique. However, when serum samples were used as antigen, the method was less sensitive (1.5-2.0 ng) because some of the serum proteins migrate with the phosphatase and decrease the intensity of the stain for acid phosphatase. For this reason we could not detect the phosphatase in serum samples of normal persons; only patients with moderately (or greater) increased activity in their serum showed positive results. In contrast, by radioimmunoassay as little as 1.0 ng of the phosphatase can be detected in serum.

1966 ◽  
Vol 12 (4) ◽  
pp. 226-233 ◽  
Author(s):  
Bernard Klein ◽  
Morris Oklander ◽  
Stanley Morgenstern

Abstract A procedure is presented for the automated determination of acid phosphatase activity in biological materials using the Robot Chemist. Although either phenylphosphate and α-naphthylphosphate may be used as substrate in this analysis, the procedure is described in detail for serum acid phosphatase using α-naphthylphosphate in 0.1 M citrate, pH 5.2, since this substrate is more selective for prostatic acid phosphatase in human serum. Enzymically generated aα- naphthol is determined by the Emerson reaction (alkaline aminoantipyrine and ferricyanide), modified for use with this automated system. Correlations are presented between the results obtained on the Robot Chemist and the identical procedure developed for the AutoAnalyzer.


1981 ◽  
Vol 27 (10) ◽  
pp. 1747-1752 ◽  
Author(s):  
G L Wright ◽  
P F Schellhammer ◽  
D N Brassil ◽  
S M Sieg ◽  
M S Leffell

Abstract We evaluated and compared five commercial radioimmunoassay kits with a standard counter-immunoelectrophoretic assay for the measurement of prostatic acid phosphatase in serum. Four of the five radioimmunoassays performed as described by the supplier with respect to sensitivity, stability, precision, linearity, analytical recovery, and expected values for the normal male population. None of the radioimmunoassays was more clinically sensitive then the counter-immunoelectrophoretic assay for detecting increased prostatic acid phosphatase in serum. Results obtained by counter-immunoelectrophoretic assay agreed with results obtained by radioimmunoassay in 96% of the tests. The proportion of positive results in patients with confirmed prostatic adenocarcinoma increased with disease progression. The fewer positive tests in localized adenocarcinoma (Stages A and B) suggests that neither the counter-immunoelectrophoretic assay nor the radioimmunoassay procedures are useful for screening unselected populations for adenocarcinoma of the prostate. The high percentage of normal values found in those patients clinically free of disease after treatment is encouraging and supports the use of the prostatic acid phosphatase immunoassays in prospectively monitoring the treatment of prostatic cancer patients.


1990 ◽  
Vol 36 (8) ◽  
pp. 1450-1455 ◽  
Author(s):  
Linh Nguyen ◽  
Alcide Chapdelaine ◽  
Simone Chevalier

Abstract We developed an assay to measure at acid pH the phosphotyrosine phosphatase activity in sera from patients with prostatic cancer. The method used quantifies the inorganic phosphate liberated from phosphotyrosine after incubation with serum, followed by the deproteinization of the reaction mixture. A high acid phosphatase (EC 3.1.3.2) activity towards phosphotyrosine was observed in all sera from patients with increased activity of prostatic acid phosphatase. This activity represented 96% of prostatic acid phosphatase and 77% of total acid phosphatase activities. Moreover, it was correlated (r = 0.91) with the amount of serum prostatic acid phosphatase determined by radioimmunoassay. When serum acid phosphatase activity was measured on several phosphorylated substrates, preferential hydrolysis was demonstrated for those in which the phosphate group was esterified on an aromatic ring rather than those presenting an aliphatic chain. Among phosphoamino acids, only phosphotyrosine was a good substrate, with little or no activity observed with phosphoserine and phosphothreonine. Human seminal plasma and partially purified prostatic acid phosphatase, tested for their activity on some of these substrates, gave similar results. On the other hand, sera from patients with above-normal alkaline phosphatase activity and no prostatic disease showed little or no activity on phosphotyrosine at both acid and alkaline pH values. Evidence is presented that the prostatic acid phosphatase in serum is a specific phosphotyrosine acid phosphatase.


1966 ◽  
Vol 12 (5) ◽  
pp. 289-298 ◽  
Author(s):  
Bernard Klein ◽  
Joseph Auerbach

Abstract α-Naphthylphosphate is a selective substrate for the differentiation of prostatic acid phosphatase from erythrocytic acid phosphatase in human serum; its comparative specificity for prostatic enzyme is demonstrated by inhibition studies and by assay of mixtures of both enzymes in serum.


Author(s):  
Hina Shamshad ◽  
Ali Sayqal ◽  
Jahan Zeb ◽  
Agha Zeeshan Mirza

Abstract A simple, accurate and precise RP-HPLC method was developed for the simultaneous determination of chloroquine, pyrimethamine and cetirizine hydrochloride concentrations in bulk drug and human serum. The assay was performed using a mobile phase of methanol: water (70:30) at pH of 2.8 ± 0.05 on the Purospher C-18 column with UV detection at 230 nm and rosuvastatin used as an internal standard. The retention times observed for chloroquine, pyrimethamine and cetirizine hydrochloride were 3.5, 2.5 and 5.5 minutes, respectively. The method was found to be specific for the assayed drugs showing a linear response in the concentration range of 1–100 μg mL−1 with coefficients of determination values of (r = 0.999). The method was developed and validated according to ICH guidelines. The method was used to monitor the serum samples and was found to be sensitive for therapeutic purposes, showing the potential to be a useful tool for routine analysis in laboratories.


2016 ◽  
Vol 129 ◽  
pp. 205-212 ◽  
Author(s):  
Adrian Marcelo Granero ◽  
Gastón Darío Pierini ◽  
Sebastián Noel Robledo ◽  
María Susana Di Nezio ◽  
Héctor Fernández ◽  
...  

2011 ◽  
Vol 6 ◽  
pp. ACI.S7346 ◽  
Author(s):  
Ani Mulyasuryani ◽  
Arie Srihardiastutie

A conductimetric enzyme biosensor for uric acid detection has been developed. The uricase, as enzyme, is isolated from Candida utilis and immobilized on a nata de coco membrane-Pt electrode. The biosensor demonstrates a linear response to urate over the concentration range 1-6 ppm and has good selectivity properties. The response is affected by the membrane thickness and pH change in the range 7.5-9.5. The response time is three minutes in aqueous solutions and in human serum samples. Application of the biosensor to the determination of uric acid in human serum gave results that compared favourably with those obtained by medical laboratory. The operational stability of the biosensor was not less than three days and the relative error is smaller than 10%.


1997 ◽  
Vol 3 (2) ◽  
pp. 47-50
Author(s):  
Walter L Strohmaier ◽  
Andreas Zumbraegel ◽  
Lennart Koschella ◽  
K Horst Bichler

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