The ability of polymers to
disrupt lipid bilayer membranes at different pH values was performed
as previously described.52 (link) Briefly, whole
blood from de-identified patients was acquired from the Vanderbilt
Technologies for Advanced Genomics (VANTAGE) core. Blood was centrifuged
to pellet erythrocytes, plasma was aspirated, erythrocytes were resuspended
in pH 7.4 PBS (Gibco), and washed three times. After the final wash,
erythrocytes were resuspended in pH 7.4, 7.0, 6.6, 6.2, or 5.8 PBS
(150 nM). Polymers were mixed with suspended erythrocytes to a concentration
of 1 μg/mL in a 96-well V-bottom plate. The plates were incubated
for 1 h at 37 °C and centrifuged to pellet intact erythrocytes,
and the supernatant was transferred to a 96-well flat-bottom plate.
Membrane disruption was quantified through hemoglobin leakage, which
can be measured using absorbance spectroscopy at 575 nm.
disrupt lipid bilayer membranes at different pH values was performed
as previously described.52 (link) Briefly, whole
blood from de-identified patients was acquired from the Vanderbilt
Technologies for Advanced Genomics (VANTAGE) core. Blood was centrifuged
to pellet erythrocytes, plasma was aspirated, erythrocytes were resuspended
in pH 7.4 PBS (Gibco), and washed three times. After the final wash,
erythrocytes were resuspended in pH 7.4, 7.0, 6.6, 6.2, or 5.8 PBS
(150 nM). Polymers were mixed with suspended erythrocytes to a concentration
of 1 μg/mL in a 96-well V-bottom plate. The plates were incubated
for 1 h at 37 °C and centrifuged to pellet intact erythrocytes,
and the supernatant was transferred to a 96-well flat-bottom plate.
Membrane disruption was quantified through hemoglobin leakage, which
can be measured using absorbance spectroscopy at 575 nm.