Jazayeri M H, Amani H, Pourfatollah A A, Avan A, Ferns G A, Pazoki-Toroudi H
Immunology Research Center, Iran University of Medical Sciences, Tehran, Iran.
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Cancer Gene Ther. 2016 Oct;23(10):365-369. doi: 10.1038/cgt.2016.42. Epub 2016 Oct 14.
Prostate-specific antigen (PSA) is used to screen for prostate disease, although it has several limitations in its application as an organ-specific or cancer-specific marker. Furthermore, a highly specific/sensitive and/or label-free identification of PSA still remains a challenge in the diagnosis of prostate anomalies. We aimed to develop a gold nanoparticle (GNP)-conjugated anti-PSA antibody-based localized surface plasmon resonance (LSPR) as a novel approach to detect prostatic disease. A total of 25 nm colloidal gold particles were prepared followed by conjugation with anti-PSA pAb (GNPs-PSA pAb). LSPR was used to monitor the absorption changes of the aggregation of the particles. The size, shape and stability of the GNP-anti-PSA were evaluated by dynamic light scattering transmission electron microscopy (TEM) and zetasizer. The GNPs-conjugated PSA-pAb was successfully synthesized and subsequently characterized using ultraviolet absorption spectroscopy and TEM to determine the size distribution, crystallinity and stability of the particles (for example, stability of GNP: 443 mV). To increase the stability of the particles, we pegylated GNPs using an N-(3-dimethylaminopropyl)-N*-ethylcarbodiimide hydrochloride (EDC)/N-hydroxylsuccinimide (NHS) linker (for example, stability of GNP after pegylation: 272 mV). We found a significant increase in the absorbance and intensity of the particles with extinction peak at 545/2 nm, which was shifted by ~1 nm after conjugation. To illustrate the potential of the GNPs-PSA pAb to bind specifically to PSA, LSPR was used. We found that the extinction peak shifted 3 nm for a solution of 100 nM unlabeled antigen. In summary, we have established a novel approach for improving the efficacy/sensitivity of PSA in the assessment of prostate disease, supporting further investigation on the diagnostic value of GNP-conjugated anti-PSA/LSPR for the detection of prostate cancer.
前列腺特异性抗原(PSA)用于筛查前列腺疾病,尽管其作为器官特异性或癌症特异性标志物在应用中有一些局限性。此外,在前列腺异常诊断中,对PSA进行高度特异性/敏感性和/或无标记识别仍然是一项挑战。我们旨在开发一种基于金纳米颗粒(GNP)偶联抗PSA抗体的局域表面等离子体共振(LSPR)技术,作为检测前列腺疾病的新方法。制备了共25nm的胶体金颗粒,然后与抗PSA多克隆抗体(GNPs-PSA pAb)偶联。LSPR用于监测颗粒聚集时的吸收变化。通过动态光散射、透射电子显微镜(TEM)和zeta电位仪评估GNP-抗PSA的尺寸、形状和稳定性。成功合成了GNPs偶联的PSA-pAb,随后使用紫外吸收光谱和TEM对其进行表征,以确定颗粒的尺寸分布、结晶度和稳定性(例如,GNP稳定性:443mV)。为了提高颗粒的稳定性,我们使用N-(3-二甲氨基丙基)-N*-乙基碳二亚胺盐酸盐(EDC)/N-羟基琥珀酰亚胺(NHS)连接剂对GNPs进行聚乙二醇化(例如,聚乙二醇化后GNP稳定性:272mV)。我们发现颗粒在545/2nm处的消光峰吸光度和强度显著增加,偶联后峰值偏移约1nm。为了说明GNPs-PSA pAb特异性结合PSA的潜力,使用了LSPR技术。我们发现,对于100nM未标记抗原溶液,消光峰偏移了3nm。总之,我们建立了一种新方法来提高PSA在前列腺疾病评估中的效能/敏感性,支持进一步研究GNP偶联抗PSA/LSPR在前列腺癌检测中的诊断价值。