A Simplified Purification Procedure for Human Prostate Antigen

Wang, Ming C.; Valenzuela, Luis; Murphy, Gerald P.; Chu, Ming

doi:10.1159/000225594

Cited by 67 publications

(33 citation statements)

References 2 publications

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“…PSA is a 33 kDa glycoprotein with chymotrypsin like activity that is produced primarily by the human prostatic epithelium and secreted into the seminal fluid [2,3]. The only well recognized physiological function of PSA is the digestion of the seminogelins and fibronectin present in seminal coagulum, liquefying the seminal clot shortly after ejaculation [4].…”

Section: Introductionmentioning

confidence: 99%

Thiophilic-interaction chromatography of enzymatically active tissue prostate-specific antigen (T-PSA) and its modulation by zinc ions

Babu

Vijayalakshmi

Smith

et al. 2008

Journal of Chromatography B

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Prostate-specific antigen (PSA) is a serine protease secreted both by normal prostate glandular epithelia cells and prostate cancer cells. We explored "thiophilic-interaction chromatography" (TIC) to isolate tissue prostate-specific antigen (T-PSA) from fresh human prostate cancer tissue harvested by radical prostatectomy for the purpose to characterize T-PSA for its enzymatic activity and sensitivity to zinc ions. We have shown, for the first time, that T-PSA has strong affinity for the thiophilic gel (T-gel). The average recovery of T-PSA from T-gel is over 87%. The presence of PSA in the column eluate was confirmed by ELISA and SDS/PAGE. Western blot developed with monoclonal antibody to PSA revealed that T-PSA was predominantly in the "free" form having a molecular weight of 33kDA. Furthermore, T-PSA was found to be enzymatically active. T-PSA was found to be less enzymatically active as compared to seminal plasma PSA. The inhibition of enzymatic activity of both f-PSA and T PSA over a wide range of concentrations of Zn 2+ ions (10 nM to 50 μM) was comparable. In contrast, the enzymatic activity of chymotrypsin, another serineprotease, was affected differently. At higher concentrations of Zn 2+ (10 μM and higher) the enzymatic activity of chymotrypsin was inhibited, whereas, at lower concentrations of Zn 2+ (5 μM and lower), the enzymatic activity was enhanced.

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Section: Introductionmentioning

confidence: 99%

Thiophilic-interaction chromatography of enzymatically active tissue prostate-specific antigen (T-PSA) and its modulation by zinc ions

Babu

Vijayalakshmi

Smith

et al. 2008

Journal of Chromatography B

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“…The changes in concentration are independent of other protein; its changes depend on serum testosterone levels [73]. If the leakage of PSA from the prostate gland, the level of PSA becomes low and high level of PSA is allied with prostatic pathology including prostatitis, BPH, and prostate cancer [74][75][76][77][78]. In 1982, PSA was used to describe as prostate cancer marker, PSA first screening report came into existence in 1991 [79,80].…”

Section: Psamentioning

confidence: 99%

Detection of Prostate Cancer: A Review

Rani

Singh

Arjuna

et al. 2018

Asian J Pharm Clin Res

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Prostate cancer is the most commonly diagnosed non-cutaneous cancer in men. Prostate-specific antigen (PSA) is the biomarker used for the screening of prostate cancer and other prostate-related problems. Not only the genetic factors are involved dietary factors, environmental factors but also responsible for the development of prostate cancer. Risk factors such as family history, age, chemical exposure, infection, and smoking are at the peak point for the development of prostate disease. Advanced age is one of the main risk factors. Radical prostatectomy is the most common therapy for small group of patients with high-grade tumors. Early screening of PSA reduces the incidence rate of prostate cancer. Mostly prostate abnormalities are seen in among male patients above the age of 50 years or older. In worldwide population, the epidemiology of prostate cancer is high in western countries and less in Asian countries.

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“…This complex procedure produced a low yield of purified PSA. A simplified purification was developed thereafter by eliminating Sephadex G-75 and preparative PAGE steps, and with an additional DEAE step, which resulted in a greater yield (3-fold) of purified PSA [4], The homogeneity of the purified PSA preparation was indicated by a single protein band with no contaminating components in sodium dodecyl sulfate (SDS)-PAGE in the presence or absence of reducing agent, and under isoelectric focusing that revealed a sin 124 Tumor Biol 1997:18:123- 134 Chu gle isoelectric point of 6.9. Furthermore, the purified PSA was shown to have a molecular weight of 33,000 by Sephadex 75 and 34,000 by SDS-PAGE with no subunit components.…”

Section: Discovery and Purification Of Psamentioning

confidence: 99%

“…A simplified purification procedure of PSA from prostate tissue and also from seminal plasma was reported by Dr. Wang in 1982 [4,14]. This improved procedure along with PSA monoclonal antibody prepared by Dr. Papsidero in 1983 [15] greatly facilitated the trans fer of our PSA technology to biomedical in dustry, which subsequently has made PSA reagents readily available since 1986 (table 1).…”

Section: Psa and Prostate Cancermentioning

confidence: 99%

Prostate-Specific Antigen and Early Detection of Prostate Cancer

Chu

1997

Tumor Biol

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Prostate-specific antigen (PSA) is biochemically a 33-kDa serine protease and is the prototype of tumor marker most useful in investigating basic science and clinical application of prostate cancer. As an immunohistopathological marker, PSA is especially effective in the identification of distant metastatic prostate carcinoma and in the differential diagnosis of poorly differentiated transitional cell carcinomas of the bladder from prostate carcinoma. As a serologic marker, PSA is most useful in staging, monitoring and in early detection of recurrent disease. The greatest value of PSA is as a screening aid along with digital rectal examination for early detection of prostate cancer. PSA-based screening tests have been performed in various regions of the world and have yielded prostate cancer detection rates proportional to that of incidence rates in the countries. Most significantly, PSA detects early nonpalpable prostate cancer. Approximately 85% of the prostate tumors detected through PSA have the clinical features associated with medically important cancer. Further, a majority of the tumors, 70%, are confined to the organ. Although the issues of cost-benefit and increase in ultimate survival rate are still being evaluated in two large-scale randomized national trials, timely and definitive therapies of these potentially lethal tumors can result in high cure rate and improve the quality of patients’ life.

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A Simplified Purification Procedure for Human Prostate Antigen

Abstract: Methods for purifying the human prostate specific antigen are described. The antigen was isolated from human prostate and seminal plasma. Purified antigen (ca. 1 mg/ml) from seminal plasma was immunologically identical and biochemically similar to that of prostatic tissue.

Cited by 67 publications

References 2 publications

Thiophilic-interaction chromatography of enzymatically active tissue prostate-specific antigen (T-PSA) and its modulation by zinc ions

Thiophilic-interaction chromatography of enzymatically active tissue prostate-specific antigen (T-PSA) and its modulation by zinc ions

Detection of Prostate Cancer: A Review

Prostate-Specific Antigen and Early Detection of Prostate Cancer

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