CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2025 , Vol. 43 >Issue 4: 497 - 502

DOI: https://doi.org/10.12140/j.issn.1000-7423.2025.04.008

ORIGINAL ARTICLES

Selection and identification of aptamers against the circulating cathodic antigen of Schistosoma mansoni

  • TANG Qi ,
  • QIU Jiayin ,
  • LI Jiajia ,
  • ZHOU Xinjie ,
  • LV Chao ,
  • FENG Ting ,
  • XU Jing ,
  • QIN Zhiqiang
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  • National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; NHC Key Laboratory on Parasite and Vector Biology; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai 200025, China

Received date: 2025-04-11

  Revised date: 2025-05-12

  Online published: 2025-10-09

Supported by

Key Discipline Projects under the Shanghai Three-Year Action Plan for Strengthening Public Health System Construction (2023-2025)(GWVI-11.1-12);National Key Research and Development Program of China(2021YFC2300800);National Key Research and Development Program of China(2021YFC2300803)

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Abstract

Objective To screen and identify the aptamer specifically targeting the circulating cathodic antigen (CCA) of Schistosoma mansoni. Methods The recombinant prokaryotic expression vector pET-28a-CCA was constructed, and the expression of pET-28a-CCA protein was induced and purified by Ni-NTA column. The expression characteristics of pET-28a-CCA were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Using pET-28a-CCA protein as the target, systematic evolution of ligands by exponential enrichment (SELEX) was performed. Quantitative real-time PCR was used to monitor and analyze each round of library enrichment. After multiple rounds of stringent selection, the obtained products were subjected to high-throughput sequencing analysis, bioinformatics analysis, and the characterization of binding affinity of candidate aptamers to the pET-28a-CCA protein using microscale thermophoresis (MST), followed by molecular docking. Results Double enzyme digestion and sequencing results confirmed that the pET-28a-CCA recombinant plasmid contained an approximately 1 044 bp CCA gene fragment. SDS-PAGE results demonstrated successful expression of the pET-28a-CCA protein, with a relative molecular mass (Mr) of approximately 43 600. The SELEX screening results showed that the library of the 13th screening round reached the best enrichment. Based on sequencing results, nine candidate aptamer sequences were selected. Among them, seven aptamers exhibited binding affinity to the pET-28a-CCA as determined by MST analysis. TQ-apt1 had the lowest Gibbs free energy (ΔG = -13.09 kcal/mol), the highest GC content (59.2%), the highest melting temperature (102.3 ℃), and the highest G-quadruplex score (15 points). TQ-apt1 had the highest affinity with pET-28a-CCA protein with a dissociation constant of 2.1 nmol/L. Conclusion In this study, aptamers targeting the pET-28a-CCA protein were successfully obtained through SELEX technology.

Key words： Schistosoma mansoni; Circulating cathodic antigen; Aptamers; Systematic evolution of ligands by exponential enrichment; Recombinant prokaryotic protein

Cite this article

TANG Qi , QIU Jiayin , LI Jiajia , ZHOU Xinjie , LV Chao , FENG Ting , XU Jing , QIN Zhiqiang . Selection and identification of aptamers against the circulating cathodic antigen of Schistosoma mansoni[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025 , 43(4) : 497 -502 . DOI: 10.12140/j.issn.1000-7423.2025.04.008

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