Glycanostics, Ltd., Bratislava, Slovak Republic.
Institute of Chemistry, Bratislava, Slovak Republic.
PLoS One. 2024 Mar 18;19(3):e0300430. doi: 10.1371/journal.pone.0300430. eCollection 2024.
The glycoprofiling of two proteins, the free form of the prostate-specific antigen (fPSA) and zinc-α-2-glycoprotein (ZA2G), was assessed to determine their suitability as prostate cancer (PCa) biomarkers. The glycoprofiling of proteins was performed by analysing changes in the glycan composition on fPSA and ZA2G using lectins (proteins that recognise glycans, i.e. complex carbohydrates). The specific glycoprofiling of the proteins was performed using magnetic beads (MBs) modified with horseradish peroxidase (HRP) and antibodies that selectively enriched fPSA or ZA2G from human serum samples. Subsequently, the antibody-captured glycoproteins were incubated on lectin-coated ELISA plates. In addition, a novel glycoprotein standard (GPS) was used to normalise the assay. The glycoprofiling of fPSA and ZA2G was performed in human serum samples obtained from men undergoing a prostate biopsy after an elevated serum PSA, and prostate cancer patients with or without prior therapy. The results are presented in the form of an ROC (Receiver Operating Curve). A DCA (Decision Curve Analysis) to evaluate the clinical performance and net benefit of fPSA glycan-based biomarkers was also performed. While the glycoprofiling of ZA2G showed little promise as a potential PCa biomarker, the glycoprofiling of fPSA would appear to have significant clinical potential. Hence, the GIA (Glycobiopsy ImmunoAssay) test integrates the glycoprofiling of fPSA (i.e. two glycan forms of fPSA). The GIA test could be used for early diagnoses of PCa (AUC = 0.83; n = 559 samples) with a potential for use in therapy-monitoring (AUC = 0.90; n = 176 samples). Moreover, the analysis of a subset of serum samples (n = 215) revealed that the GIA test (AUC = 0.81) outperformed the PHI (Prostate Health Index) test (AUC = 0.69) in discriminating between men with prostate cancer and those with benign serum PSA elevation.
评估了两种蛋白质(游离前列腺特异性抗原(fPSA)和锌-α-2-糖蛋白(ZA2G))的糖基化特征,以确定它们作为前列腺癌(PCa)生物标志物的适用性。使用凝集素(识别聚糖的蛋白质,即复杂碳水化合物)分析 fPSA 和 ZA2G 上糖基组成的变化来进行蛋白质的糖基化特征分析。使用辣根过氧化物酶(HRP)和选择性地从人血清样本中富集 fPSA 或 ZA2G 的抗体修饰的磁性珠(MB)进行蛋白质的特异性糖基化特征分析。随后,将抗体捕获的糖蛋白孵育在凝集素包被的 ELISA 板上。此外,还使用了新型糖蛋白标准(GPS)来对测定进行归一化。在接受前列腺活检的血清 PSA 升高的男性和接受过或未接受过治疗的前列腺癌患者的人血清样本中进行了 fPSA 和 ZA2G 的糖基化特征分析。结果以 ROC(接收器操作曲线)的形式呈现。还进行了 DCA(决策曲线分析)以评估 fPSA 糖基化生物标志物的临床性能和净收益。虽然 ZA2G 的糖基化特征分析作为潜在的 PCa 生物标志物的前景不大,但 fPSA 的糖基化特征分析似乎具有重要的临床潜力。因此,GIA(糖基生物活检免疫测定)试验整合了 fPSA 的糖基化特征分析(即 fPSA 的两种糖型)。GIA 试验可用于 PCa 的早期诊断(AUC = 0.83;n = 559 个样本),并具有用于治疗监测的潜力(AUC = 0.90;n = 176 个样本)。此外,对一组血清样本(n = 215)的分析表明,在区分前列腺癌男性和良性血清 PSA 升高的男性方面,GIA 试验(AUC = 0.81)优于 PHI(前列腺健康指数)试验(AUC = 0.69)。