The pH-MUC1 NP solution was diluted with three solutions of PBS buffer at pH 7.4, 6.8, and 5.5, respectively, to achieve a final concentration of 1.6 mg pH-MUC1 NP/mL. Next, 10 μL of fluorescamine solution in DMSO (3 mg/mL) was mixed with 150 μL of each NP solution. Each solution was then transferred into a black flat bottom 96-well plate and analyzed using a plate reader (Spectra Max M3 by Molecular devices). PBS solutions at pH 7.4, 6.8, and 5.5 and PBS at pH 7.4, 6.8, and 5.5 with pH-MUC1 NPs without fluorescamine were used as controls.
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