Citation: Yaliang Huang, Ting Sun, Wendi Li, Lin Liu, Gang Liu, Xinyao Yi, Jianxiu Wang. Colorimetric and electrochemical detection of ligase through ligation reaction-induced streptavidin assembly[J]. Chinese Chemical Letters, ;2022, 33(6): 3151-3155. doi: 10.1016/j.cclet.2021.10.038 shu

Colorimetric and electrochemical detection of ligase through ligation reaction-induced streptavidin assembly

Yaliang Huang ^{a, b} ,
Ting Sun ^b ,
Wendi Li ^c ,
Lin Liu ^{b, *,} ,
Gang Liu ^b ,
Xinyao Yi ^a ,
Jianxiu Wang ^{a, *,}

a.
Hunan Provincial Key Laboratory of Micro & Nano Materials Interface Science, College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, China
b.
Henan Province of Key Laboratory of New Optoelectronic Functional Materials, Anyang Normal University, Anyang 455000, China
c.
Sanquan College of Xinxiang Medical University, Xinxiang 453003, China

liulin@aynu.edu.cn

jxiuwang@csu.edu.cn

Received Date: 29 July 2021
Revised Date: 5 October 2021
Accepted Date: 14 October 2021
Available Online: 22 October 2021

Figures(5)

We propose a concept for ligase detection by conversion of aggregation-based homogeneous analysis into surface-tethered electrochemical assay through streptavidin (SA)-biotin interaction. Sortase A (SrtA) served as the model analyte and two biotinylated peptides (bio-LPETGG and GGGK-bio) were used as the substrates. SrtA-catalyzed ligation of the peptide substrates led to the generation of bio-LPETGGGK-bio. The ligation product (bio-LPETGGGK-bio) induced the aggregation and color change of SA-modified gold nanoparticles (AuNPs) through the SA-biotin interactions, which could be assayed by the colorimetric method. Furthermore, we found that the bio-LPETGGGK-bio could trigger the assembly of tetrameric SA proteins with the formation of the (SA-bio-LPETGGGK-bio)_n assemblies through the same interactions. The above results were further confirmed by atomic force microscopy and fluorescent imaging. The insulated assemblies were in-situ fabricated at the SA-modified gold electrode, thus hindering the electron transfer of [Fe(CN)₆]^3−/4− and leading to an increase in the electron-transfer resistance. The capability of the method for the detection of SrtA both in vitro and Staphylococcus aureus (S. aureus) has been demonstrated. SrtA with a concentration down to 1 pmol/L has been determined by the electrochemical analysis, which is lower than that achieved by the colorimetric assay (50 pmol/L). By integrating the advantages of homogeneous reaction and heterogeneous detection, the strategy serves as an ideal means for the fabrication of various sensing platforms by adopting biotin-labeled and sequence-specific peptide or nucleic acid substrates.
- Ligase,
- Electrochemical analysis,
- Streptavidin,
- Gold nanoparticles,
- Colorimetric assay,
- Electrochemical impedance spectroscopy
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