scholarly journals On Chip Immuno-Affinity Profiling of Cancer- and Benign Hyperplasia-Associated Free Prostate Specific Antigen

2011 ◽  
Vol 31 (2) ◽  
pp. 111-118 ◽  
Author(s):  
Maja M. Kosanovic ◽  
Sanja R. Goc ◽  
Goran S. Potpara ◽  
Miroslava M. Jankovic
Keyword(s):  
Molecular Mass ◽  
Prostate Specific Antigen ◽  
Seminal Plasma ◽  
Specific Antigen ◽  
Analytical Techniques ◽  
Molecular Forms ◽  
Striking Difference ◽  
Free Psa ◽  
Lower Molecular Mass ◽  
Benign Hyperplasia

Prostate specific antigen (PSA) exhibits pronounced heterogeneity in both primary structure and glycan composition, resulting in the existence of different molecular forms. Investigation of PSA structure is a demanding task facing limitations due to inadequate sensitivity of analytical techniques and low concentrations of the different forms. This study aimed to profile free PSA (fPSA), especially lower molecular mass species lacking detailed classification, in normal seminal plasma and in sera from subjects with benign hyperplasia (BPH) or cancer of the prostate (PCa) as samples of known clinical relevance. fPSA forms were separated from complex proteomes on chips with immobilized anti-fPSA antibody followed by detection using surface-enhanced laser desorption/ionization time of flight mass spectrometry. At least 39 fPSA-immunoreactive species, ranging from 3–29 kDa were detected in seminal plasma. General fPSA profiles in seminal plasma and sera were similar, but differed in the abundance and presence of particular peaks/clusters of the lower molecular mass species. No striking difference in fPSA forms was observed between BPH and PCa samples, but some distinct peaks varied in intensity and frequency within or between groups. Obtained data verify fPSA heterogeneity that might be important for better exploration of all their molecular and marker potentials.

scholarly journals Structural Diversity of Cancer-related and Non-Cancer-related Prostate-specific Antigen

2002 ◽  
Vol 48 (12) ◽  
pp. 2187-2194 ◽  
Author(s):  
Takahiro Isono ◽  
Tsutomu Tanaka ◽  
Susumu Kageyama ◽  
Tatsuhiro Yoshiki
Keyword(s):  
Prostate Cancer ◽  
Molecular Mass ◽  
Prostate Specific Antigen ◽  
Seminal Fluid ◽  
Polyclonal Antibodies ◽  
Specific Antigen ◽  
Structural Diversity ◽  
Molecular Forms ◽  
Critical Importance ◽  
Lower Molecular Mass

Abstract Background: Heterogeneity among the various molecular forms of prostate-specific antigen (PSA) has not been well characterized, despite the critical importance of PSA in the detection of prostate cancer. The purpose of this study was to examine PSA heterogeneity in cancerous and noncancerous materials by extensive and systematic protein analysis. Methods: A catalog of molecular forms of PSA was established with the PSA purified from seminal fluid. This catalog was used to analyze PSA heterogeneity in cancerous and noncancerous materials by immunoblotting with polyclonal antibodies. Results: PSA from noncancerous materials showed a wider range of molecular mass, from 6000 to 28 000 Da. PSA from cancerous materials did not contain lower molecular mass forms. Conclusions: The PSA protein catalog may be useful for the analysis of differences among PSA forms in men with and without prostate cancer and for analysis of antibodies used to detect PSA.

Purification and characterization of prostate-specific antigen (PSA) complexed to alpha 1-antichymotrypsin: potential reference material for international standardization of PSA immunoassays

1995 ◽  
Vol 41 (9) ◽  
pp. 1273-1282 ◽  
Author(s):  
Z Chen ◽  
A Prestigiacomo ◽  
T A Stamey
Keyword(s):  
Molecular Mass ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Molar Ratio ◽  
Free Psa ◽  
Apparent Homogeneity ◽  
International Standardization

Abstract We describe for the first time a protocol to purify to apparent homogeneity an in vitro-prepared complex of prostate-specific antigen (PSA) and alpha 1-antichymotrypsin (ACT) by using a combination of gel filtration and ion-exchange chromatography. The purity of the PSA-ACT complex was confirmed by gel electrophoresis and Western blot. The PSA-ACT complex was stable in the pH range 6.0 to 7.8; it was also stable in various matrices, temperatures, and high concentrations of salt. Purification of the PSA-ACT complex was highly reproducible. An absorptivity of 0.99 L x g-1 x cm-1 at 280 nm was assigned to the PSA-ACT complex, based on amino acid analysis. Because PSA and ACT bind in a 1:1 molar ratio, we determined the molecular mass of the PSA-ACT complex as the mass encoded by the cDNA of ACT (plus 26% carbohydrate) plus the molecular mass of PSA (28,430 Da), which totals 89,280 Da. Using this material, we made two common calibrators, one of 100% PSA-ACT complex and one of 90% PSA-ACT complex plus 10% free PSA by volume (90:10 calibrator). Substitution of these calibrators for the manufacturers' calibrators in nine commercial immunoassays substantially reduced differences between immunoassays, especially for serum PSA values between 4 and 10 micrograms/L. The 90:10 calibrator is recommended as a universal calibrator for international standardization of PSA immunoassays.

scholarly journals Characterization of Novel Monoclonal Antibodies for Prostate-Specific Antigen (PSA) with Potency to Recognize PSA Bound to α2-Macroglobulin

2005 ◽  
Vol 51 (1) ◽  
pp. 84-92 ◽  
Author(s):  
Yvonne Baumgart ◽  
Andreas Otto ◽  
Angelika Schäfer ◽  
Elke Usbeck ◽  
Christiane Cott ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Molecular Forms ◽  
Phage Display Technology ◽  
Free Psa ◽  
Α2 Macroglobulin ◽  
Display Technology

Abstract Background: Different molecular forms of prostate-specific antigen (PSA) have been used to differentiate between benign prostatic hyperplasia and prostate cancer. Detecting PSA bound to endogenous inhibitors such as α1-antichymotrypsin (ACT) and α2-macroglobulin (α2M) is often difficult because of epitope masking or sensitivity problems. Here we report the characterization of four novel mouse monoclonal antibodies (mabs) obtained by immunization with PSA-α2M complexes. Their ability to detect free PSA and PSA-inhibitor complexes was shown, and their epitopes were analyzed by phage display technology. Methods: The properties of the mabs were studied by competition and sandwich assays and by Western blotting. Epitope mapping was performed by screening of a phage display peptide library. Results: All four mabs recognized free PSA, PSA-ACT, and PSA-α2M complexes, but to various degrees. With different combinations of mabs in competition experiments, antibodies were identified that enhance binding of other mabs to PSA, forming the molecular basis of a very sensitive assay for the detection of PSA and PSA-ACT complexes. Mabs with highest reactivity for PSA-α2M were selected to establish an immunoassay for that complex. Western blot analysis revealed that all mabs recognized conformational epitopes of PSA. These findings were supported by phage display results demonstrating mimotopes in the PSA molecule. Conclusion: The results presented here could aid in the further development of clinically relevant assays for PSA and PSA-α2M complexes.

scholarly journals Human Serum Low Molecular Mass Prostate-Specific Antigen as Biomarker

2017 ◽  
Vol 36 (4) ◽  
pp. 322-330
Author(s):  
Sanja Goč ◽  
Miroslava Janković
Keyword(s):  
Molecular Mass ◽  
Prostate Specific Antigen ◽  
Rare Species ◽  
Specific Antigen ◽  
Immunoaffinity Chromatography ◽  
Striking Difference ◽  
Individual Species ◽  
Profile Characteristic ◽  
Pathophysiological Condition ◽  
On Chip

SummaryBackground: Prostate-specific antigen (PSA) is a glycoprotein tumor marker known to exist as numerous glycospecies. Investigations on its glycobiochemical properties aimed at their use in the preparation of adjuncts in determining PSA concentration for clinical purposes have accumulated a lot of data on its structural properties. In this study, we reconsidered unexplored ubiquitously present low molecular mass species of PSA regarding to molecular mass, origin and pathophysiological source specificity in order to evaluate them as biomarkers. Methods: Data on low molecular mass PSA-immunoreactive species from sera of subjects with prostate cancer (PCa), benign prostatic hyperplasia (BPH), breast cancer (BCa), and urine of healthy males obtained by on-chip immunoaffinity chromatography combined with mass spectrometry were analyzed. Results: The results obtained indicated PSA species common to BCa, PCa, and BPH at 12-13 kDa, 17-19 kDa and 21-24 kDa. The striking difference in predominant frequencies made the profile characteristic in each examined pathophysiological condition. On the other hand, paired groups of prostatic and extraprostatic PSA contained rare species with small differences among groups concerning individual species. Low molecular mass PSA also included rare species unique for each group of samples. Conclusion: The results obtained revealed that uniformity of low molecular mass PSA-immunoreactive species in sera prevails over diversity related to cancer and non-cancer conditions, but at the same time some of them are molecules with biomarker potential for BPH detection.

Effect of Marathon Running on Total and Free Serum Prostate-Specific Antigen Concentrations

2003 ◽  
Vol 127 (3) ◽  
pp. 345-348 ◽  
Author(s):  
Alexander Kratz ◽  
Kent B. Lewandrowski ◽  
Arthur J. Siegel ◽  
Patrick M. Sluss ◽  
Kelly Y. Chun ◽  
...  
Keyword(s):  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Physical Exertion ◽  
The United States ◽  
Marathon Running ◽  
Reliable Determination ◽  
Free Psa ◽  
Sporting Event ◽  
Exercise Induced ◽  
Total Psa

Abstract Context.—Prostate-specific antigen (PSA) is an important tumor marker for the most frequently diagnosed cancer in the United States. A major limitation of this marker is falsely elevated results in patients who are found not to have prostate cancer. The effects of vigorous physical exertion on PSA concentrations are controversial. Objective.—To determine the effects of marathon running on PSA levels. Design.—Measurement of total and free PSA levels in the sera of participants in a marathon before and within 4 and 24 hours after the race. Results.—None of the participants had elevated total PSA levels before the race. Although we found no statistically significant changes in average total or free PSA concentrations at either time point, after the marathon, 2 (11%) of 18 runners had total PSA concentrations outside the standard reference range. Changes in total PSA levels did not correlate with age or prerace PSA concentrations. Free PSA levels were not statistically significantly changed after the race and did not allow a reliable determination of exercise-induced PSA elevations. Conclusions.—Although it may not be necessary for men to abstain from exercise involving running before blood draws for PSA analysis, elevated PSA concentrations may be observed in some individuals after participation in a major sporting event. In these cases, repeat measurements should be considered at a time significantly removed from such exercise.

scholarly journals The Combination of Human Glandular Kallikrein and Free Prostate-specific Antigen (PSA) Enhances Discrimination Between Prostate Cancer and Benign Prostatic Hyperplasia in Patients with Moderately Increased Total PSA

1999 ◽  
Vol 45 (11) ◽  
pp. 1960-1966 ◽  
Author(s):  
Angeliki Magklara ◽  
Andreas Scorilas ◽  
William J Catalona ◽  
Eleftherios P Diamandis
Keyword(s):  
Prostate Cancer ◽  
Benign Prostatic Hyperplasia ◽  
Prostate Specific Antigen ◽  
Prostatic Carcinoma ◽  
Specific Antigen ◽  
Area Under The Curve ◽  
Prostatic Hyperplasia ◽  
Free Psa ◽  
Glandular Kallikrein ◽  
Total Psa

Abstract Background: Prostate-specific antigen (PSA) is the most reliable tumor marker available and is widely used for the diagnosis and management of prostate cancer. Unfortunately, PSA cannot distinguish efficiently between benign and malignant disease of the prostate, especially within the range of 4–10 μg/L. Among the refinements developed to enhance PSA specificity is the free/total PSA ratio, which is useful in discriminating between the two diseases within the diagnostic “gray zone”. Recent data indicate that human glandular kallikrein (hK2), a protein with high homology to PSA, may be an additional serum marker for the diagnosis and monitoring of prostate cancer. Methods: We analyzed 206 serum samples (all before treatment was initiated) from men with histologically confirmed benign prostatic hyperplasia (n = 100) or prostatic carcinoma (n = 106) with total PSA in the range of 2.5–10 μg/L. Total and free PSA and hK2 were measured with noncompetitive immunological procedures. Statistical analysis was performed to investigate the potential utility of the various markers or their combinations in discriminating between benign prostatic hyperplasia and prostatic carcinoma. Results: hK2 concentrations were not statistically different between the two groups of patients. There was a strong positive correlation between hK2 and free PSA in the whole patient population. hK2/free PSA ratio (area under the curve = 0.69) was stronger predictor of prostate cancer than the free/total PSA ratio (area under the curve = 0.64). At 95% specificity, the hK2/free PSA ratio identified 30% of patients with total PSA between 2.5–10 μg/L who had cancer. At 95% specificity, the hK2/free PSA ratio identified 25% of patients with total PSA between 2.5 and 4.5 μg/L who had cancer. Conclusions: Our data suggest that hK2 in combination with free and total PSA can enhance the biochemical detection of prostate cancer in patients with moderately increased total PSA concentrations. More specifically, the hK2/free PSA ratio appears to be valuable in identifying a subset of patients with total PSA between 2.5 and 4.5 μg/L who have high probability of cancer and who should be considered for biopsy.

A microfluidic immunoassay platform for the detection of free prostate specific antigen: a systematic and quantitative approach

The Analyst ◽  
2015 ◽  
Vol 140 (13) ◽  
pp. 4423-4433 ◽  
Author(s):  
Narayanan Madaboosi ◽  
Ruben R. G. Soares ◽  
Virginia Chu ◽  
João Pedro Conde
Keyword(s):  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Quantitative Approach ◽  
Free Psa ◽  
Microfluidic Immunoassay ◽  
Antigen A ◽  
Relevant Range

A novel physisorption- and bio-affinity amplification-based microfluidic immunoassay platform for free PSA detection within a clinically relevant range is reported.

scholarly journals High-throughput glycopeptide profiling of prostate-specific antigen from seminal plasma by MALDI-MS

Talanta ◽  
2021 ◽  
Vol 222 ◽  
pp. 121495
Author(s):  
Wei Wang ◽  
Anna Kałuża ◽  
Jan Nouta ◽  
Simone Nicolardi ◽  
Mirosława Ferens-Sieczkowska ◽  
...  
Keyword(s):  
Prostate Specific Antigen ◽  
High Throughput ◽  
Seminal Plasma ◽  
Specific Antigen ◽  
Maldi Ms

scholarly journals Molecular Forms of Prostate-specific Antigen in Malignant and Benign Prostatic Tissue: Biochemical and Diagnostic Implications

2000 ◽  
Vol 46 (1) ◽  
pp. 47-54 ◽  
Author(s):  
Klaus Jung ◽  
Brigitte Brux ◽  
Michael Lein ◽  
Birgit Rudolph ◽  
Glen Kristiansen ◽  
...  
Keyword(s):  
Prostate Specific Antigen ◽  
Specific Antigen ◽  
Gel Filtration ◽  
Tumor Stage ◽  
Prostatic Hyperplasia ◽  
Prostatic Tissue ◽  
Molecular Forms ◽  
Tissue Samples ◽  
Total Psa ◽  
Lower Ratio

Abstract Background: Patients with prostate cancer (PCa) show a lower ratio of free prostate-specific antigen (fPSA) to total PSA (tPSA) in serum than patients with benign prostatic hyperplasia (BPH). The patterns of the intracellular PSA isoforms in malignant and benign prostatic tissue have been studied as potential molecular reasons for this phenomenon. Methods: Prostatic tissue samples were obtained after cystoprostatectomy from patients with bladder cancer (n = 10), from BPH patients (transurethral resection of the prostate, n = 10; adenomectomy, n = 10), and from the cancerous and noncancerous parts of the same prostates removed surgically by prostatectomy because of PCa (n = 20). PSA pattern was characterized by gel filtration, immunoblotting, and immunoassays for tPSA, fPSA, α1-antichymotrypsin-PSA (ACT-PSA), and complexed PSA (Bayer Immuno 1 assay). Comparisons were made with the PSA concentrations in serum. Results: The major portion of tPSA in all tissue samples was fPSA; complexed PSA forms were <2%. Samples from cystoprostatectomy patients had the lowest and those from adenomectomy patients the highest values of tPSA and fPSA. PSA concentrations were lower in cancerous than in the noncancerous parts of the prostate. No significant correlations were found between tumor stage or grade and the amounts of tPSA, fPSA, and ACT-PSA in tissue. Tissue PSA values were not correlated with the serum PSA concentrations nor with the ratios fPSA/tPSA and ACT-PSA/tPSA in sera. Conclusions: The amounts of tPSA and the PSA isoforms in prostatic tissue explain neither the concentrations of tPSA and PSA isoforms in serum nor the behavior of the ratio fPSA/tPSA in patients with BPH and PCa.

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