An amphiphilic conjugate based on Ce6, luminol, and PEG (abbreviated as CLP) was synthesized by conjugating both luminol and PEG with Ce6 using a standard N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS)–activated condensation reaction. Briefly, 50 mg (0.08 mmol) of Ce6 was dissolved in 10 ml of anhydrous DMSO, into which EDC [236 mg (1.23 mmol)] and NHS [144 mg (1.25 mmol)] were sequentially added. The reaction mixture was stirred at 50°C in the dark for 17 hours. Then, luminol [28 mg (0.16 mmol)] was added to the activated solution. After 3 days, monomethyl-terminated PEG monoamine with a molecular weight of 2000 [320 mg (0.16 mmol)] was added. Reaction was conducted at 50°C in the dark for 4 days. Last, the reaction mixture was dialyzed (molecular weight cutoff, 3500) against deionized water to remove unreacted reagents and by-products. After filtration through a 0.22-μm syringe filter, the obtained aqueous solution was lyophilized to give rise to a dark green powder.

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