Electrochemiluminescence Distance and Reactivity of Coreactants Determine the Sensitivity of Bead-Based Immunoassays

Yafeng Wang; Jialian Ding; Ping Zhou; Jilin Liu; Zhiyuan Qiao; Kai Yu; Jie Jiang; Bin Su

doi:10.1002/anie.202216525

Electrochemiluminescence Distance and Reactivity of Coreactants Determine the Sensitivity of Bead-Based Immunoassays

Angew Chem Int Ed Engl. 2023 Apr 11;62(16):e202216525. doi: 10.1002/anie.202216525. Epub 2023 Mar 8.

Authors

Yafeng Wang¹, Jialian Ding¹, Ping Zhou¹, Jilin Liu², Zhiyuan Qiao³, Kai Yu³, Jie Jiang^{2

3

4}, Bin Su¹

Affiliations

¹ Key Laboratory of Excited-State Materials of Zhejiang Province, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
² School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, 150001, China.
³ School of Environment, School of Marine Science and Technology (Weihai), Harbin Institute of Technology, Weihai, 150090, China.
⁴ State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin, 150090, China.

PMID: 36812044
DOI: 10.1002/anie.202216525

Abstract

Herein we report the study of electrochemiluminescence (ECL) generation by tris(2,2'-bipyridyl)ruthenium (Ru(bpy)₃ ²⁺ ) and five tertiary amine coreactants. The ECL distance and lifetime of coreactant radical cations were measured by ECL self-interference spectroscopy. And the reactivity of coreactants was quantitatively evaluated in terms of integrated ECL intensity. By statistical analysis of ECL images of single Ru(bpy)₃ ²⁺ -labeled microbeads, we propose that ECL distance and reactivity of coreactant codetermine the emission intensity and thus the sensitivity of immunoassay. 2,2-bis(hydroxymethyl)-2,2',2''-nitrilotriethanol (BIS-TRIS) can well balance ECL distance-reactivity trade-off and enhance the sensitivity by 236 % compared with tri-n-propylamine (TPrA) in the bead-based immunoassay of carcinoembryonic antigen. The study brings an insightful understanding of ECL generation in bead-based immunoassay and a way of maximizing the analytical sensitivity from the aspect of coreactant.

Keywords: Electrochemiluminescence; Immunoassay; Radical Lifetime; Reaction Mechanism; Reactivity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

2,2'-Dipyridyl* / chemistry
Luminescent Measurements / methods
Microspheres
Ruthenium* / chemistry

Substances

2,2'-Dipyridyl
Ruthenium