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10 mg/mL of mPEG-PEAD and heparin were separately prepared in PBS and sterilized using a 0.22 µm filter membrane. Then, 2 μL of AAL (0.5 mg/mL) was first mixed with 10 μL of heparin solution. Thereafter, 80 μL of mPEG-PEAD solution was added to the heparin:AAL mixture. The final mass ratio was 1:100:800 of AAL:heparin:mPEG-PEAD. An increase in the solution turbidity was immediately observed due to the instantaneous formation of Coa. Particle size of fabricated AAL loaded mPEG-Coa or empty mPEG-Coa (i.e., without AAL loading) was measured by Malvern Zetasizer.
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