Multi-Enzyme Cascade Nanoreactors for High-Throughput Immunoassay: Transitioning Concept in Lab to Application in Community

Anal Chem. 2024 Jul 16;96(28):11463-11471. doi: 10.1021/acs.analchem.4c01912. Epub 2024 Jul 4.

Abstract

In this work, we reported a cholesterol oxidase (Chox)-loaded platinum (Pt) nanozyme with the collaborative cascade nanoreactor for the construction of nanozyme-enzyme-linked immunosorbent assay (N-ELSA) models to realize high-throughput rapid evaluation of cancer markers. Considering the high specific surface area and manipulable surface sites, ZIF-8 was used as a substrate for natural enzyme and nanozyme loading. The constructed ZIF-8-Pt nanozyme platform exhibited efficient enzyme-like catalytic efficiency with a standard corrected activity of 60.59 U mg-1, which was 12 times higher than that of the ZIF-8 precursor, and highly efficient photothermal conversion efficiency (∼35.49%). In N-ELISA testing, developed multienzyme photothermal probes were immobilized in microplates based on antigen-antibody-specific reactions. Cholesterol was reacted in a cascade to reactive oxygen radicals, which attacked 3,3',5,5'-tetramethylbenzidine, causing it to oxidize and color change, thus exhibiting highly enhanced efficient photothermal properties. Systematic temperature evaluations were performed by a hand-held microelectromechanical system thermal imager under the excitation of an 808 nm surface light source to determine the cancer antigen 15-3 (CA15-3) profiles in the samples. Encouragingly, the temperature signal from the microwells increased with increasing CA15-3, with a linear range of 2 mU mL-1 to 100 U mL-1, considering it to be the sensor with the widest working range for visualization and portability available. This work provides new horizons for the development of efficient multienzyme portable colorimetric-photothermal platforms to help advance the community-based process of early cancer detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzidines / chemistry
  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / metabolism
  • Cholesterol / analysis
  • Cholesterol / chemistry
  • Cholesterol / metabolism
  • Cholesterol Oxidase* / chemistry
  • Cholesterol Oxidase* / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • High-Throughput Screening Assays
  • Humans
  • Platinum* / chemistry
  • Zeolites / chemistry

Substances

  • Platinum
  • Cholesterol Oxidase
  • 3,3',5,5'-tetramethylbenzidine
  • Biomarkers, Tumor
  • Benzidines
  • Cholesterol
  • Zeolites