Sensitive colorimetric assay of T4 DNA ligase by the oxidase nanozyme of LaMnO3.26 coupled with a hyperbranched amplification reaction

Abstract

The development of efficient methods for the detection of T4 DNA ligase is extremely important for public health. The present work demonstrates the integration of engineerable oxidase nanozyme of LaMnO_3.26 nanomaterials for the colorimetric detection of T4 DNA ligase. Specifically, the LaMnO_3.26 nanomaterials exhibited oxidase-like activity, oxidizing o-phenylenediamine (OPD), 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), and 3,3′,5,5′-tetramethylbenzidine (TMB) to their corresponding oxidation products, which featured maximum absorption wavelengths at 450, 417 and 650 nm, respectively, while pyrophosphate ion (PPi) caused an obvious decrease in the oxidase-like activity of LaMnO_3.26 through its surface coordination with the surface-exposed Mn element and induced aggregation of the nanozyme. Attributed to the PPi regulated oxidase nanozyme activity, LaMnO_3.26 served as a colorimetric probe for the quantitative detection of T4 DNA ligase assisted by a hyperbranched amplification reaction for signal amplification. The T4 DNA ligase was detected with a linear range of 4.8 × 10⁻³ to 6.0 U mL⁻¹, achieving a detection limit of 1.6 × 10⁻³ U mL⁻¹. The outcome indicated that the developed nanozyme might be extended to a broad range of practical applications.