Aptamer-Mediated Enrichment of Rare Circulating Fetal Nucleated Red Blood Cells for Noninvasive Prenatal Diagnosis

Abstract

Isolation of circulating fetal nucleated red blood cells (cfNRBCs) from maternal peripheral blood provides a superior strategy for noninvasive prenatal genetic diagnosis. Recent technical advances in single-cell isolation and genetic analyses have promoted the clinical application of circulating fetal cell-based noninvasive prenatal diagnosis. However, the lack of highly specific ligands for rare circulating fetal cell enrichment from massive maternal cells significantly impedes the clinical transformation progress. In this work, aptamers specific to NRBCs were developed through clinical sample-based cell-SELEX. Herein, the complex clinical system provides natural selection stringency through binding competition between target and background cells, and it empowers aptamers with high specificity. An aptamer-based strategy was also established to isolate cfNRBCs from maternal peripheral blood. Results show the remarkable selectivity and affinity of developed aptamers, enabling efficient enrichment of cfNRBCs from abundant maternal cells. Moreover, screening for fetal sex and trisomy syndrome achieved high accuracy through chromosome analysis of enriched cfNRBCs. To the best of our knowledge, this is the first report to develop aptamer ligands for cfNRBC enrichment, providing an efficient strategy to screen cfNRBC-specific ligands and demonstrating broad application potential for cfNRBC-based noninvasive prenatal diagnosis.

The Development and Characterisation of ssDNA Aptamers via a Modified Cell-SELEX Methodology for the Detection of Human Red Blood Cells

“…In this study, we successfully selected novel ssDNA aptamers that target human red blood cells, a vital target analyte within forensic analysis and an analyte of interest in a medical setting. Aptamers have previously been developed for a range of surface proteins of the RBC, such as Glycophorin A, the Rhesus D antigenic epitope and A1 antigen, as well as RBCs of medical interest, such as rare circulating foetal RBCs and malaria parasiteinfected RBCs [58][59][60][61][62]. However, to the best of our knowledge, this is the first reported use of a modified Cell-SELEX methodology [22] that has used the entire healthy isolated RBC from a pool of donors as the selection target, with negative selection included to increase the stringency of selection.…”

Leveraging Aptamer-Based DNA Nanotechnology for Bioanalysis and Cancer Therapeutics

“…7 For example, by modifying on the surface of functional materials, the aptamer LXD-11b selected from cell-SELEX can specifically recognize nucleated red blood cells in maternal peripheral blood. 8 In this Account, we discuss the integration of DNA aptamers with DNA nanotechnology, focusing on our studies in molecular medicine, specifically in bioanalysis and cancer therapeutics. The first part delves into aptamers for cancer bioanalysis, covering molecular modifications, aptamer-tethered DNA nanostructures, and aptamer-involved molecular computation.…”

“…On the other hand, triggered by aptamers, molecular computation based on dynamic DNA nanotechnology can be employed for intelligent bioanalysis and clinical diagnosis . For example, by modifying on the surface of functional materials, the aptamer LXD-11b selected from cell-SELEX can specifically recognize nucleated red blood cells in maternal peripheral blood …”

Aptamer-Based Multiparameter Analysis for Molecular Profiling of Hematological Malignancies

“…Aptamers, also known as chemical antibodies, can be obtained by systematic evolution of ligands by exponential enrichment against whole living cells (Cell-SELEX), even without prior knowledge of molecular signatures on the cell surface. − Meanwhile, taking advantage of simple synthesis, convenient modification, and high chemical stability, aptamers represent powerful molecular recognition tools for precise cell typing. − In this study, a panel of aptamers able to discriminate subtle surface protein differences was acquired through aptamer selection against K562 and multidrug-resistant K562 (K562/D) cell lines. Moreover, a machine learning model was developed for multivariate data statistical analysis of the aptamer binding panels, and the potential of these selected aptamers for molecular typing of HM was evaluated through multiparameter mapping and applied to the classification of complex clinical samples (Figure ).…”