Table 1. Characteristic Properties of the HRP-Encapsulating LUV₂₀₀ Used in This Study.

property	fractions 16	overall vesicle fraction a
[POPC]	2.17 mM	1.10 mM
n(POPC)	1.69 μmol	≈6.0 μmol
mean hydrodynamic vesicle diameter	180 nm	180 nm
POPC molecules per vesicle (NPOPC)b	≈2.78 × 105	≈2.78 × 105
volume of one vesicle (Vves)	2.88 × 10−18 L	2.88 × 10−18 L
[vesicle]c	7.8 nM	4.0 nM
number of vesicles (Nves)d	3.66 × 1012	1.30 × 1013
[HRP]overall	92 nM	37 nM
n(HRP)	72 pmol	200 pmol
number of HRP molecules per vesicle (NHRP)e	≈12	≈9
[HRP]inside f	6.8 μM	5.3 μM

^aFractions 15–21 were considered. Each fraction had a volume of 0.78 mL; total volume of all vesicle fractions, 5.46 mL. Amounts applied onto the Sepharose 4B column, ≈6.0 μmol POPC (= 0.5 mL 12 μmol mL⁻¹) and ≈16 nmol HRP (= 0.5 mL 31 nmol mL⁻¹) (Figure 5).

^bCalculated by assuming monodisperse, unilamellar and spherical vesicles. Further, a thickness of the POPC bilayer of 3.7 nm and a mean POPC headgroup area of 0.72 nm² were taken into account.⁶¹,⁶²

^c[vesicle] = [POPC]/N_POPC.

^dN_ves = n(POPC)N_A/N_POPC; N_A = 6.02 × 10²³, Avogadro’s number.

^eN_HRP = n(HRP)N_A/N_ves = [HRP]_overall/[vesicle].

^f[HRP]_inside = n(HRP)/(V_vesN_ves) = N_HRP/(V_vesN_A).