Heterodimeric aptamer improves the limit of detection of sandwich ELASA and dot blot assay

IF 2.5 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 2025-09-25 DOI:10.1016/j.ab.2025.115984

Tzi Shien Yeoh, Marimuthu Citartan

引用次数: 0

Abstract

Heterodimerization, which is an amalgamation of discrete aptamers into a single functional module is a viable strategy to augment the binding strength of aptamers. In this study, we sought to derive several heterodimeric aptamers using the discrete aptamers, LepDapt-2a and LepDapt-5a prior to their binding evaluation against the target LipL32. We further integrated heterodimeric aptamer LepDapt-H1b as a detection aptamer into aptamer-based sandwich ELASA and dot blot assay for the detection of Leptospira, attaining a LOD of 4 and 10 CFU/mL for both assays, respectively, which are better compared to our previous study. Additionally, the LOQ acquired for sandwich ELASA assay was 11 CFU/mL. We have proven that heterodimerization has improved the performance of sandwich ELASA and dot blot assay, suggesting them to be applicable for the diagnostics of leptospirosis.

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异二聚体提高了夹心ELASA和斑点印迹法的检出限。

异二聚化是将离散的适体合并成一个单一的功能模块，是增强适体结合强度的一种可行策略。在这项研究中，我们试图在LepDapt-2a和LepDapt-5a与目标LipL32结合评估之前，使用离散的适体推导出几种异二聚体适体。我们进一步将LepDapt-H1b作为检测适配体整合到基于适配体的三明治式ELASA法和点印迹法中检测钩端螺旋体，两种检测方法的LOD分别为4和10 CFU/mL，与我们之前的研究相比有更好的提高。此外，夹心ELASA法获得的定量限为11 CFU/mL。我们已经证明，异源二聚化提高了夹心ELASA和斑点印迹法的性能，表明它们适用于钩端螺旋体病的诊断。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.