Diamandis E P, Nadkarni S, Bhaumik B, Abdelrahman A, Melegos D N, Borchert G, Black M H, Alonso M, Salas A, de los Toyos J R, Sampedro A, López-Otín C
Department of Pathology, Mount Sinai Hospital, Canada.
Clin Chem. 1997 Aug;43(8 Pt 1):1365-71.
We developed mouse monoclonal antibodies (Abs) against pepsinogen C with highly purified antigen isolated from gastric mucosa. The Abs were used to construct a two-site sandwich-type assay for pepsinogen C with time-resolved fluorometry as a detection technique. The assay has a detection limit of 0.1 microgram/L and is precise (within-run and day-to-day CVs < 11%). We used this assay to measure pepsinogen C in seminal plasma, breast cyst fluid, amniotic fluid, male and female serum, serum from patients with prostate cancer, urine, breast tumor cytosolic extracts, breast milk, and cerebrospinal fluid. Highest pepsinogen C concentrations were in seminal plasma, followed by breast cyst fluid and amniotic fluid. We found no correlation between prostate-specific antigen concentrations and concentrations of pepsinogen C in serum of prostate cancer patients, and concluded that this marker is not useful for either diagnosing or monitoring prostatic carcinoma. The availability of a highly sensitive, reliable, and convenient method for quantifying pepsinogen C will allow investigations into the possible diagnostic value of this analyte in various clinical conditions, including benign breast diseases, breast cancer, fertility, and pregnancy.
我们利用从胃黏膜中分离出的高纯度抗原，研制出了抗胃蛋白酶原C的小鼠单克隆抗体。这些抗体被用于构建一种采用时间分辨荧光法作为检测技术的胃蛋白酶原C双位点夹心型检测方法。该检测方法的检测限为0.1微克/升，且精密度高（批内和批间变异系数<11%）。我们使用此检测方法对精浆、乳腺囊肿液、羊水、男性和女性血清、前列腺癌患者血清、尿液、乳腺肿瘤胞质提取物、母乳和脑脊液中的胃蛋白酶原C进行了检测。胃蛋白酶原C浓度最高的是精浆，其次是乳腺囊肿液和羊水。我们发现前列腺癌患者血清中前列腺特异性抗原浓度与胃蛋白酶原C浓度之间无相关性，并得出结论，该标志物对前列腺癌的诊断或监测均无用处。一种用于定量胃蛋白酶原C的高灵敏度、可靠且便捷的方法的出现，将有助于研究该分析物在包括良性乳腺疾病、乳腺癌、生育能力和妊娠等各种临床状况下可能的诊断价值。
