Alkaline Phosphatase-Triggered Simultaneous Hydrogelation and Chemiluminescence

Area of Science:

• Biomaterials Science
• Chemical Biology
• Analytical Chemistry

Background:

• Chemiluminescence (CL) offers superior signal-to-noise ratios compared to fluorescence.
• Enzyme-instructed self-assembly (EISA) is a crucial process in biomaterials and drug delivery.
• Direct tracking of EISA using CL has not been previously reported.

Purpose of the Study:

• To develop and validate a novel chemiluminescence (CL) method for directly characterizing and imaging enzyme-instructed self-assembly (EISA).
• To investigate the use of CL to monitor the simultaneous hydrogelation process triggered by alkaline phosphatase (ALP).

Main Methods:

• Coincubation of a hydrogelator precursor (Fmoc-Phe-Phe-Tyr(H2PO3)-OH, 1P) with a CL agent (AMPPD, 2) and alkaline phosphatase (ALP).
• Systematic study of 1P hydrogelation with and without 2 under ALP catalysis.
• Analysis of CL properties of 2 with and without 1P under ALP catalysis.
• Time-course CL imaging using an IVIS optical imaging system.

Main Results:

• The CL agent AMPPD (2) was identified as an ideal indicator for ALP-triggered hydrogelation of 1P.
• Simultaneous EISA and hydrogelation of 1P were successfully tracked and imaged in real-time using CL.
• The CL method demonstrated high sensitivity and signal-to-noise ratio for monitoring the EISA process.

Conclusions:

• Chemiluminescence provides a powerful and direct method for characterizing and imaging enzyme-instructed self-assembly (EISA) processes.
• This CL-based approach enables real-time monitoring of hydrogel formation, with potential applications in tracking biological EISA events.