Serum prostate-specific acid phosphatase: development and validation of a specific radioimmunoassay.

1978 ◽  
Vol 24 (11) ◽  
pp. 1915-1919 ◽  
Author(s):  
P Vihko ◽  
E Sajanti ◽  
O Jänne ◽  
L Peltonen ◽  
R Vihko
Keyword(s):  
Acid Phosphatase ◽  
Reference Group ◽  
Prostatic Acid Phosphatase ◽  
Human Spleen ◽  
Specific Radioimmunoassay ◽  
Chloramine T ◽  
Monospecific Antisera ◽  
Synovial Tissues ◽  
Development And Validation ◽  
Free Serum

Abstract We describe radioimmunoassay for human prostatic acid phosphatase [orthophosphoric-monoester phospho-hydrolase (acid optimum), EC 3.1.3.2] in serum, with use of monospecific antisera raised in rabbits against highly purified acid phosphatase from human prostates. The antiserum did not cross react with partly purified acid phosphatases from human spleen, erythrocytes, or synovial tissues. 125I-labeled acid phosphatase was prepared by a Chloramine T method, and the bound and free antigen was separated in the assay by use of anti-rabbit gamma-globulin raised in sheep. Uniform low nonspecific binding of the [125I]acid phosphatase was achieved by using acid-phosphatase-free serum to prepare standard curves and diluted samples of serum with high acid phosphatase activities. Concentrations of immunoreactive acid phosphatase in the serum of healthy men ranged from less than 1 to 10 microgram/liter and for 12 patients with advanced prostatic carcinoma between 100 and 500 microgram/liter. The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group.

Rapid radioimmunoassay for prostate-specific acid phosphatase in human serum.

1980 ◽  
Vol 26 (11) ◽  
pp. 1544-1547 ◽  
Author(s):  
P Vihko ◽  
A Kostama ◽  
O Jänne ◽  
E Sajanti ◽  
R Vihko
Keyword(s):  
Acid Phosphatase ◽  
Benign Prostatic Hyperplasia ◽  
Prostatic Carcinoma ◽  
Reference Group ◽  
Prostatic Acid Phosphatase ◽  
Prostatic Hyperplasia ◽  
Serum Concentrations ◽  
Healthy Men ◽  
Monospecific Antisera ◽  
Group Serum

Abstract We describe a rapid radioimmunoassay for human prostatic acid phosphatase (EC 3.1.3.2) in serum, with use of monospecific antisera raised in rabbits against the primary highly purified acid phosphatase (pl 4.9) from human prostates, and with a second antibody-polyethylene glycol porecipitation. This radioimmunoassay is sensitive and can be performed within 5 h. Concentrations of the immunoreactive acid phosphatase in sera of healthy men (n = 394) ranged from 0.3 to 3.6 microgram/L (mean 1.94, SD 0.66 microgram/L). Concentrations of the enzyme in sera of men with benign prostatic hyperplasia (n = 56) or with carcinoma of nonprostatic origin (n = 24) were identical with those of the reference group. Serum concentrations of immunoreactive prostatic acid phosphatase of patients with occult, non-metastatic, and metastatic prostatic carcinoma varied from 1.7 to 9.3 (n = 9), 4.2 to 59.4 (n = 12), and 20 to 198 (n = 10) microgram/L, respectively. The amount of immunoassayable prostatic acid phosphatase was unchanged for at least five days in serum stored at 4 degrees C.

scholarly journals A critical evaluation of a specific radioimmunoassay for prostatic acid phosphatase

Cancer ◽  
1982 ◽  
Vol 50 (9) ◽  
pp. 1847-1851 ◽  
Author(s):  
S. Larry Goldenberg ◽  
Hulbert K. B. Silver ◽  
Lorne D. Sullivan ◽  
Michael J. Morse ◽  
E. L. Archibald
Keyword(s):  
Acid Phosphatase ◽  
Critical Evaluation ◽  
Prostatic Acid Phosphatase ◽  
Specific Radioimmunoassay

Comparison of Bolton-Hunter and Chloramine-T Techniques for the Radioiodination of Prostatic Acid Phosphatase

1983 ◽  
Vol 20 (2) ◽  
pp. 112-115 ◽  
Author(s):  
C M Sturgeon ◽  
L Beynon ◽  
J Seth ◽  
G D Chisholm
Keyword(s):  
Acid Phosphatase ◽  
Detection Limit ◽  
Prostatic Acid Phosphatase ◽  
Chromatographic Purification ◽  
Double Antibody ◽  
Chloramine T

A double antibody, semi-automated radioimmunoassay for serum prostatic acid phosphatase (PAP) is described, which uses the 125I-labelled N-succinimidyl-3-(4-hydroxyphenyl) propionate ester of PAP. This type of label has substantially higher immunoreactivity than that prepared using the chloramine-T method of radioiodination. Chromatographic purification of either label on Ultrogel ACA44 further improved immunoreactivity. The lowest detection limit (0·35 μg/l) was achieved with the chromatographically purified ester label.

scholarly journals Development and Validation of a Specific Radioimmunoassay for Somatostatin in Human Plasma

1979 ◽  
Vol 16 (1-6) ◽  
pp. 15-25 ◽  
Author(s):  
Erica Penman ◽  
J. A. H. Wass ◽  
Alison Lund ◽  
P. J. Lowry ◽  
Jennifer Stewart ◽  
...  
Keyword(s):  
Coefficient Of Variation ◽  
Normal Subjects ◽  
Plasma Binding ◽  
Fasting Level ◽  
Specific Radioimmunoassay ◽  
Chloramine T ◽  
The Mean ◽  
Development And Validation ◽  
Circulating Levels ◽  
Free Plasma

Little is known about the factors controlling somatostatin secretion in man, and data are not available on the changes in circulating levels in various human physiological or pathophysiological states. This is mainly a consequence of the technical difficulties involved in measuring somatostatin in plasma. In the presence of plasma, binding of somatostatin tracer to antibody was consistently decreased by about 20%, and this could not be abolished by the addition of EDTA and aprotinin or by the use of specially prepared somatostatin-free plasma. Furthermore, in the presence of plasma, endogenous somatostatin does not dilute in parallel with synthetic cyclic somatostatin standard. We have, therefore, developed and validated a radioimmunoassay for somatostatin using prior extraction of the peptide onto leached silica glass. Tyrosine-11 somatostatin was iodinated using lactoperoxidase and purified on ODS silica. This method is superior to iodination using chloramine-T with CMC cellulose purification, and gives a highly purified preparation with a shelf-life of at least eight weeks. Using this tracer and a specific antiserum, the limit of sensitivity of the assay was 10 pg/ml, with an intra-assay coefficient of variation of 12% (n = 16) and inter-assay coefficient of variation of 15% (n = 10). Parallelism has been demonstrated between standard synthetic cyclic somatostatin and all extracted plasma samples. The mean recovery of exogenous somatostatin from plasma was 78%. The fasting level of immunoreactive somatostatin at 0900 hours in 40 normal subjects ranged from 17 to 81 pg/ml. Care is needed, however, when comparing these values with those obtained from other laboratories since standard preparations of somatostatin vary considerably in their immunopotency.

Rapid, fully automated radioimmunoassay of prostatic acid phosphatase in serum.

1980 ◽  
Vol 26 (11) ◽  
pp. 1583-1587 ◽  
Author(s):  
S Dass ◽  
N L Bowen ◽  
K D Bagshawe
Keyword(s):  
Acid Phosphatase ◽  
Prostatic Acid Phosphatase ◽  
Metastatic Carcinoma ◽  
Control Group ◽  
Control Groups ◽  
Carcinoma Of The Prostate ◽  
Lower Detection Limit ◽  
Lower Detection ◽  
Specific Antisera ◽  
Chloramine T

Abstract Prostatic acid phosphatase was purified from human semen and its purity established by biochemical and immunological criteria. Rabbits were injected with the purified isoenzyme to raise specific antisera. The prostatic acid phosphatase was radiolabeled with 125I by the Chloramine T method. We developed a fully automated double-antibody radiommunosassay for measuring prostatic acid phosphatase in serum from patients with carcinoma of the prostate and from several control groups. The lower detection limit of the radioimmunoassay was 2.0 microgram of prostatic acid phosphatase per litre of serum. Values for most members of the control group was <2.0 microgram/L; patients with metastatic carcinoma of the prostate had values ranging from <2.0 to 300 microgram/L of serum.

Production of monoclonal antibodies against prostatic acid phosphatase by in vitro immunization of human spleen cells

1985 ◽  
Vol 84 (1-2) ◽  
pp. 105-116 ◽  
Author(s):  
Noboru Yamaura ◽  
Masanao Makino ◽  
Linda J. Walsh ◽  
Andrew W. Bruce ◽  
Byung-Kil Choe
Keyword(s):  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Prostatic Acid Phosphatase ◽  
Spleen Cells ◽  
Human Spleen ◽  
In Vitro Immunization

The Demonstration of Human Prostatic Acid Phosphatase (Pap) With Phosphorylcholine

1976 ◽  
Vol 34 ◽  
pp. 88-89
Author(s):  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Prostate Gland ◽  
Prostatic Acid Phosphatase ◽  
Human Prostate ◽  
Rat Tissues ◽  
Specific Substrate ◽  
Acid Phosphatases ◽  
Lysosomal Acid ◽  
Cacodylate Buffer

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.

Prostatic Acid Phosphatase (PAP) Differentiated Ultracytochemically from Lysosomal Acid Phosphate in the Rat with a PAP/Specific Substrate, Phosphorylcholine

1975 ◽  
Vol 33 ◽  
pp. 450-451
Author(s):  
W. Allen Shannon ◽  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Phosphate Buffer ◽  
Prostatic Carcinoma ◽  
Prostatic Acid Phosphatase ◽  
Chemotherapeutic Agents ◽  
Specific Substrate ◽  
Acid Phosphate ◽  
Lysosomal Acid ◽  
Design And Synthesis ◽  
New Chemotherapeutic Agents

During the design and synthesis of new chemotherapeutic agents for prostatic carcinoma based on phosphorylated agents which might be enzyme-activated to cytotoxicity, phosphorylcholine, [(CH3)3+NCH2CH2OPO3Ca]Cl-, has been indicated to be a very specific substrate for prostatic acid phosphatase (PAP). This phenomenon has led to the development of specific histochemical and ultracytochemical methods for PAP using modifications of the Gomori lead method for acid phosphatase. Comparative histochemical results in prostate and kidney of the rat have been published earlier with phosphorylcholine (PC) and β-glycerophosphate (βGP). We now report the ultracytochemical results.Minced tissues were fixed in 3% glutaraldehyde-0.1 M phosphate buffered (pH 7.4) for 1.5 hr and rinsed overnight in several changes of 0.05 M phosphate buffer (pH 7.0) containing 7.5% sucrose. Tissues were incubated 30 min to 2 hr in Gomori acid phosphatase medium (2) containing 0.1 M substrate, either PC or βGP.

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