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Ultrasensitive and specific fluorescence detection of a cancer biomarker via nanomolar binding to a guanidinium-modified calixarene.

ABSTRACT: We designed a water-soluble guanidinium-modified calix[5]arene to target lysophosphatidic acid (LPA), an ideal biomarker for early diagnosis of ovarian and other gynecologic cancers, achieving binding on the nanomolar level. An indicator displacement assay, coupled with differential sensing, enabled ultrasensitive and specific detection of LPA. Moreover, we show that using a calibration line, the LPA concentration in untreated serum can be quantified in the biologically relevant low ?M range with a detection limit of 1.7 ?M. The reported approach is feasible for diagnosing ovarian and other gynecologic cancers, particularly at their early stages.

SUBMITTER: Zheng Z

PROVIDER: S-EPMC5892409 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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Ultrasensitive and specific fluorescence detection of a cancer biomarker <i>via</i> nanomolar binding to a guanidinium-modified calixarene.

Zheng Zhe Z Geng Wen-Chao WC Gao Jie J Wang Yu-Ying YY Sun Hongwei H Guo Dong-Sheng DS

Chemical science 20180109 8

We designed a water-soluble guanidinium-modified calix[5]arene to target lysophosphatidic acid (LPA), an ideal biomarker for early diagnosis of ovarian and other gynecologic cancers, achieving binding on the nanomolar level. An indicator displacement assay, coupled with differential sensing, enabled ultrasensitive and specific detection of LPA. Moreover, we show that using a calibration line, the LPA concentration in untreated serum can be quantified in the biologically relevant low μM range wit ...[more]

PMID: 29675249

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Project description:Short single-stranded oligonucleotides represent a class of promising therapeutics with diverse application areas. Antisense oligonucleotides, for example, can interfere with various processes involved in mRNA processing through complementary base pairing. Also RNA interference can be regulated by antagomirs, single-stranded siRNA and single-stranded microRNA mimics. The increased susceptibility to nucleolytic degradation of unpaired RNAs can be counteracted by chemical modification of the sugar phosphate backbone. In order to understand the dynamics of such single-stranded RNAs, we investigated their fate after exposure to cellular environment by several fluorescence spectroscopy techniques. First, we elucidated the degradation of four differently modified, dual-dye labeled short RNA oligonucleotides in HeLa cell extracts by fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy and Förster resonance energy transfer. We observed that the integrity of the oligonucleotide sequence correlates with the extent of chemical modifications. Furthermore, the data showed that nucleolytic degradation can only be distinguished from unspecific effects like aggregation, association with cellular proteins, or intramolecular dynamics when considering multiple measurement and analysis approaches. We also investigated the localization and integrity of the four modified oligonucleotides in cultured HeLa cells using fluorescence lifetime imaging microscopy. No intracellular accumulation could be observed for unmodified oligonucleotides, while completely stabilized oligonucleotides showed strong accumulation within HeLa cells with no changes in fluorescence lifetime over 24 h. The integrity and accumulation of partly modified oligonucleotides was in accordance with their extent of modification. In highly fluorescent cells, the oligonucleotides were transported to the nucleus. The lifetime of the RNA in the cells could be explained by a balance between release of the oligonucleotides from endosomes, degradation by RNases and subsequent depletion from the cells.

Dataset Information

Ultrasensitive and specific fluorescence detection of a cancer biomarker via nanomolar binding to a guanidinium-modified calixarene.

Publications

Ultrasensitive and specific fluorescence detection of a cancer biomarker <i>via</i> nanomolar binding to a guanidinium-modified calixarene.

Similar Datasets

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