High-performance liquid chromatography method for detecting prostate-specific membrane antigen activity

Kamga, Isabelle Mawabo; Ng, Rudy J.; Hosaka, Mia; Berkman, Clifford E.

doi:10.1016/s0003-2697(02)00284-1

Cited by 2 publications

(2 citation statements)

References 8 publications

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“…The third approach relies on the hydrolysis of a suitable (nonnatural) GCPII substrate with the subsequent detection of reaction products by high-performance liquid chromatography (HPLC). 17–20 Although this approach offers several advantageous features (nonhazardous, sensitive, low false-positive rate), it is more suitable to low-throughput applications dealing with a limited number of samples.…”

Section: Introductionmentioning

confidence: 99%

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

et al. 2012

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Glutamate carboxypeptidase II (GCPII) is an important target for therapeutic and diagnostic interventions aimed at prostate cancer and neurologic disorders. Here we describe the development and optimization of a high-throughput screening (HTS) assay based on fluorescence polarization (FP) that facilitates the identification of novel scaffolds inhibiting GCPII. First, we designed and synthesized a fluorescence probe based on a urea-based inhibitory scaffold covalently linked to a Bodipy TMR fluorophore (TMRGlu). Next, we established and optimized conditions suitable for HTS and evaluated the assay robustness by testing the influence of a variety of physicochemical parameters (e.g., pH, temperature, time) and additives. Using known GCPII inhibitors, the FP assay was shown to be comparable to benchmark assays established in the field. Finally, we evaluated the FP assay by HTS of a 20 000–compound library. The novel assay presented here is robust, highly reproducible (Z′ = 0.82), inexpensive, and suitable for automation, thus providing an excellent platform for HTS of small-molecule libraries targeting GCPII.

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Section: Introductionmentioning

confidence: 99%

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

et al. 2012

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“…据文献报导, PSMA 可以水解 N-乙酰天冬氨酰谷氨酸, 生成谷氨酸盐和 N-乙酰天冬氨酸 [7] . 根据此原理建立的检测 PSMA 的方法主要有高效液相色谱法 [8] . 该方法灵敏度高, 但也具有前处理繁琐、费时等缺点.…”

Section: 早期诊断、基因治疗、预后评估中所起的作用变得越来越重要unclassified

Colorimetric Sensing of Prostate Specific Membrane Antigen Based on Gold Nanoparticles

Feng¹,

Gao²,

Wang³

2019

Acta Chim. Sinica

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Cancer is a major cause of death and its early diagnosis has been a research goal for many decades. For males, prostatic carcinoma has become the second leading cause of cancer death worldwide. Prostate specific membrane antigen (PSMA) has been widely recognized as a prostate cancer marker. Thus, measurement of PSMA would be more valuable for the early diagnosis of prostate cancer. Nanomaterials have the characteristics of small size effect, quantum size effect, macroscopic quantum tunneling effect and surface effect, and have been widely used in various fields, such as cell imaging, analysis and detection, drug release and treatment. Gold nanoparticles have been widely used in biosensing and medical diagnosis due to their simple preparation, high stability and unique photoelectric properties. In this paper, a new colorimetric approach is proposed for simple detection of PSMA based on gold nanoparticles. In the experiment, we synthesized gold nanoparticles with positive charges, and the polyanionic peptide as the substrate of PSMA. The detection of PSMA was based on the property that different aggregation states of gold nanoparticles can lead to the change of color and the specific recognition of PSMA for its substrate. The positively charged gold nanoparticles interact electrostatically with polyanionic peptide, resulting in aggregation of gold nanoparticles. In the presence of PSMA, however, the polyanionic peptide are hydrolyzed into glutamic acid fragment due to the reaction between the PSMA and the polyanionic peptide, resulting in the dispersion of gold nanoparticles. This behaviour leads to the development of a rapid and simple colorimetric method for assaying PSMA activity, with a detection limit of 0.5 nmol/L and the linear range of 2～10 nmol/L. This approach is simple compared to the existing ones since the gold nanoparticles-peptide based sensor is easy to be assembled and the detection can be achieved without the involvement of complicated procedures. Moreover, the applicability of the method has been demonstrated by detecting PSMA spiked into urine samples.

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High-performance liquid chromatography method for detecting prostate-specific membrane antigen activity

Cited by 2 publications

References 8 publications

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

Colorimetric Sensing of Prostate Specific Membrane Antigen Based on Gold Nanoparticles

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