Chemiluminescent duplex analysis using phenoxy-1,2-dioxetane luminophores with color modulation

Area of Science:

• Analytical Chemistry
• Biochemistry
• Diagnostic Sciences

Background:

• Multiplex technology allows simultaneous detection of multiple analytes in a single sample.
• Chemiluminescent phenoxy-dioxetane luminophores are key components in diagnostic assays.
• Predicting light emission spectra is crucial for developing multicolor assays.

Purpose of the Study:

• To design and synthesize a library of chemiluminescent dioxetane luminophores with multicolor emission wavelengths.
• To develop a chemiluminescent duplex system for simultaneous detection of two enzyme activities.
• To demonstrate the utility of this system in both physiological solutions and bacterial assays.

Main Methods:

• Designed a library of chemiluminescent dioxetane luminophores based on fluorescence-emission spectrum prediction.
• Selected two dioxetane luminophores with distinct emission spectra and similar quantum yields.
• Developed turn-ON chemiluminescent probes by equipping luminophores with enzymatic substrates.
• Performed simultaneous detection of enzyme activities using different filter slits.

Main Results:

• Successfully synthesized a library of dioxetane luminophores with multicolor emission wavelengths.
• Developed a chemiluminescent duplex system capable of simultaneously detecting two different enzymatic activities.
• Demonstrated simultaneous enzyme activity detection in physiological solution and a bacterial assay.
• Achieved distinct detection using blue and red filter slits for the two enzymes.

Conclusions:

• This work presents the first successful demonstration of a chemiluminescent duplex system using two-color phenoxy-1,2-dioxetane luminophores.
• The developed library of dioxetanes is beneficial for creating chemiluminescence luminophores for multiplex analysis.
• This technology holds promise for advancing multiplex analysis of enzymes and bioanalytes in diagnostics.