Incubation of MDCO-216 (ApoA-IMilano/POPC) with Human Serum Potentiates ABCA1-Mediated Cholesterol Efflux Capacity, Generates New Prebeta-1 HDL, and Causes an Increase in HDL Size
Table 1
Prebeta-1 HDL as measured by ELISA in MDCO-216 solutions mixed with buffer or plasma after 1 h incubation at 0 or 37°C. MDCO-216 stock solution contained 13 mg/mL ApoA-I Milano dimer, 6% sucrose, 1% mannitol, and phosphate buffer pH 7.4. Vehicle solution contained 6% sucrose, and 1% mannitol in phosphate buffer, pH 7.4. Solution High (10 mg/mL protein) was prepared by mixing 200 μL MDCO-216 stock with 60 μL Vehicle. Solution Medium (3 mg/mL protein) was prepared by mixing 100 μL MDCO-216 stock with 330 μL Vehicle. Solution Low (1 mg/mL protein) was prepared by mixing 50 μL Solution High with 450 μL Vehicle. 50 μL of these Solutions was spiked into 450 μL Vehicle (Table 1(a)) or into 450 μL plasma (Table 1(b)) and incubated 60 minutes at 0°C or 37°C.
(a)
Solution spiked
Vehicle
Low
Medium
High
Final MDCO-216 concentration (μg/mL protein):
0
100
300
1000
Prebeta-1 HDL in μg/mL (% of added)
Incubated at 0°C
0
12 (12%)
44 (15%)
118 (12%)
Incubated at 37°C
0
5 (5%)
13 (4%)
52 (5%)
(b)
Incubation temp.
1 h 0°C
Solution spiked
Vehicle
Low
Med
High
Final MDCO-216 concentration (μg/mL protein):
0
100
300
1000
Prebeta-1 HDL net due to plasma
Spiked in plasma 1
62
75
101
105
Spiked in plasma 2
126
126
131
121
Spiked in plasma 3
112
144
143
125
Spiked in plasma average
100
115
125
117
% of plasma spiked with vehicle
115%
125%
117%
Incubation temp.
1 h 37°C
Solution spiked
Vehicle
Low
Med
High
Final MDCO-216 concentration (μg/mL protein):
0
100
300
1000
Prebeta-1 HDL net due to plasma
Spiked in plasma 1
5
77
162
379
Spiked in plasma 2
12
89
153
375
Spiked in plasma 3
23
127
173
401
Spiked in plasma average
13
98
163
385
% of plasma spiked with vehicle
733%
1220%
2888%
Value for MDCO-216 spiked in plasma minus value for MDCO-216 spiked in Vehicle (Table 1(a)) in μg/mL. (value for plasma plus MDCO-216 minus value for MDCO-216 in vehicle)/value for plasma spiked with vehicle.
