Publication Date
2-1-2021
Journal
Biochimie
DOI
10.1016/j.biochi.2020.11.018
PMID
33242496
PMCID
PMC7863625
PubMedCentral® Posted Date
2-1-2022
PubMedCentral® Full Text Version
Author MSS
Published Open-Access
yes
Keywords
Aptamers, Nucleotide, Carcinoma, Pancreatic Ductal, Cell Line, Tumor, Drug Delivery Systems, Humans, Neoplasm Proteins, Pancreatic Neoplasms, Thy-1 Antigens
Abstract
Modified DNA aptamers incorporated with amino-acid like side chains or drug-like ligands can offer unique advantages and enhance specificity as affinity ligands. Thy-1 membrane glycoprotein (THY1 or CD90) was previously identified as a biomarker candidate of neovasculature in pancreatic ductal adenocarcinoma (PDAC). The current study developed and evaluated modified DNA X-aptamers targeting THY1 in PDAC. The expression and glycosylation of THY1 in PDAC tumor tissues were assessed using immunohistochemistry and quantitative proteomics. Bead-based X-aptamer library that contains 108 different sequences was used to screen for high affinity THY1 X-aptamers. The sequences of the X-aptamers were analyzed with the next-generation sequencing. The affinities of the selected X-aptamers to THY1 were quantitatively evaluated with flow cytometry. Three high affinity THY1 X-aptamers, including XA-B217, XA-B216 and XA-A9, were selected after library screening and affinity binding evaluation. These three X-aptamers demonstrated a high binding affinity and specificity to THY1 protein and the THY1 expressing cell lines, using THY1 antibody as a comparison. The development of these X-aptamers provides highly specific and non-immunogenic affinity ligands for THY1 binding in the context of biomarker development and clinical applications. They could be further exploited to assist molecular imaging of PDAC targeting THY1.
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