6.4 Membrane experiments
6.4.4 Liposome aggregation/fusion testing
LUVs were tested for aggregation and fusion by measuring turbidity using the same liposomes and buffers as used for the leakage experiments. For each experiment, liposomes were added to a solution of host in buffer such that cavitand:POPC ratio was 1:50, POPC concentration was 400 µM, DMSO total volume was 10 µL, and total sample volume was 2 mL. Turbidity was measured by measuring absorbance at 400 nm. Absorbance was recorded before LUVs were added to give 0% turbidity. After LUVs were added, each sample was stirred at 400 rpm for 30 s, then absorbance recorded continuously for 5 min with no stirring. The absorbance of LUVs with no host was measured separately to give 100% turbidity. Turbidity of calculated according to the equation
% turbidity = A(t)−Ab AL−Ab
×100 (12)
whereA(t)is the absorbance of the sample as a function of time, Ab is the absorbance of buffer, andALis the absorbance of LUVs only. Results reported are the average of two runs.
7
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8
Appendix
8.1
Documentation of copyright approval for reprinted figures
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