Toward sensitive, quantitative point-of-care testing (POCT) of protein markers: miniaturization of a homogeneous time-resolved fluoroimmunoassay for prostate-specific antigen detection

The Analyst ◽  
2011 ◽  
Vol 136 (5) ◽  
pp. 1029-1035 ◽  
Author(s):  
Annette Kupstat ◽  
Michael U. Kumke ◽  
Niko Hildebrandt
2001 ◽  
Vol 47 (4) ◽  
pp. 703-711 ◽  
Author(s):  
Timo Piironen ◽  
Martti Nurmi ◽  
Kerttu Irjala ◽  
Olli Heinonen ◽  
Hans Lilja ◽  
...  

Abstract Background: The purpose of this study was to validate the use of whole-blood samples in the determination of circulating forms of prostate-specific antigen (PSA). Methods: Blood samples of hospitalized prostate cancer and benign prostatic hyperplasia patients were collected and processed to generate whole-blood and serum samples. Three different rapid two-site immunoassays were developed to measure the concentrations of total PSA (PSA-T), free PSA (PSA-F), and PSA-α1-antichymotrypsin complex (PSA-ACT) to detect in vitro changes in whole-blood samples immediately after venipuncture. The possible influence of muscle movement on the release of PSA from prostate gland was studied in healthy men by measuring the rapid in vitro whole-blood kinetics of PSA forms before and after 15 min of physical exercise on a stationary bicycle. Results: Rapid PSA-T, PSA-F, and PSA-ACT assays were designed using a 10-min sample incubation. No significant changes were detected in the concentrations of PSA-T, PSA-F, and PSA-ACT from the earliest time point of 12–16 min compared with measurements performed up to 4 h after venipuncture. Physical exercise did not influence the concentrations of the circulating forms of PSA. Hematocrit-corrected whole-blood values of PSA-T and PSA-F forms were comparable to the respective serum values. Calculation of the percentage of PSA-F (PSA F/T ratio × 100) was similar irrespective of the sample format used, i.e., whole blood or serum. Conclusions: We found that immunodetectable PSA forms are likely at steady state immediately after venipuncture, thus enabling the use of anticoagulated whole-blood samples in near-patient settings for point-of-care testing, whereas determinations of PSA (e.g., PSA-T, PSA-F, or PSA-ACT) performed within the time frame of the office visit would provide results equivalent to conventional analyses performed in serum.


2015 ◽  
Vol 891 ◽  
pp. 120-129 ◽  
Author(s):  
Gizem Ertürk ◽  
Martin Hedström ◽  
M. Aşkın Tümer ◽  
Adil Denizli ◽  
Bo Mattiasson

2017 ◽  
Vol 98 ◽  
pp. 285-291 ◽  
Author(s):  
Bo Tian ◽  
Erik Wetterskog ◽  
Zhen Qiu ◽  
Teresa Zardán Gómez de la Torre ◽  
Marco Donolato ◽  
...  

1995 ◽  
Vol 41 (1) ◽  
pp. 54-58 ◽  
Author(s):  
H Yu ◽  
E P Diamandis

Abstract Prostate-specific antigen (PSA) is believed to be a highly specific marker for normal and cancerous prostatic tissue. We recently found that 30-40% of breast tumors produce PSA. Other data from our group suggest that normal breast can also produce PSA under conditions of stimulation by steroid hormones. In addition, we detected PSA in amniotic fluid. Here we report the presence of PSA in breast milk of lactating women. PSA concentrations in breast milk were quite variable, ranging from < 0.01 microgram/L in 4 of 38 milks to 350 micrograms/L; the median was 0.47 microgram/L. PSA concentration in breast milk was not correlated with mother's age or the sex of the newborn. It did tend to decrease with increasing time postdelivery, but was still detectable 2 weeks postdelivery. PSA in milk was equally measurable by a highly sensitive PSA assay based on time-resolved fluorometry and by the IMx automated PSA method. As confirmed by Western blot analysis, PSA in milk was present predominantly in its 33-kDa form; the PSA-alpha 1-antichymotrypsin complex (100 kDa) was also present but its concentration was < 25% of total PSA. We conclude that the female breast can produce PSA and that PSA is secreted into the milk during lactation; however, the biological role of PSA in milk is unknown. These and other data presented by our group suggest that PSA, a serine protease, may play a role in control of growth in mammary and other tissues through regulation of growth factors, cytokines, and growth-factor-binding proteins.


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