Specific Binding/Internalization of Targeted Liposomes and J591 Antibody by Cell Monolayers
| J591-labeled liposomes | A10 PSMA aptamer–labeled liposomes | J591 antibody | ||||||
| Incubation time (h) | Cell line | Liposome or antibody:receptor ratio (Liposome or antibody:cell ratio) | % specifically bound and internalized liposomes | % specifically internalized liposomes (% internalized relative to bound liposomes) | % specifically bound and internalized liposomes | % specifically internalized liposomes (% internalized relative to bound liposomes) | % specifically bound and internalized antibody | % specifically internalized antibody (% internalized relative to bound antibody) |
| 6 | LnCaP | 1:1 (18 × 104:1) | 13.2 ± 1.4 | 3.6 ± 0.9 (27.2 ± 6.7) | 7.2 ± 0.9 | 2.2 ± 0.7 (30.8 ± 10.5) | 80.4 ± 6.3 | 30.6 ± 2.9 (38.1 ± 4.7) |
| 1:10 (18 × 103:1) | 21.0 ± 2.8 | 5.5 ± 2.2 (26.2 ± 9.1) | 10.2 ± 1.2 | 3.1 ± 1.1 (30.8 ± 12.1) | 77.5 ± 3.3 | 42.7 ± 2.3 (33.5 ± 3.6) | ||
| Mat-Lu | 1:1 (9.6 × 104:1) | 6.1 ± 1.1 | 1.9 ± 0.8 (30.6 ± 15.3) | 5.4 ± 1.3 | 2.0 ± 0.1 (37.7 ± 8.3) | 78.4 ± 3.1 | 26.3 ± 2.7 (33.5 ± 3.6) | |
| 1:10 (9.6 × 103:1) | 7.6 ± 2.3 | 2.7 ± 2.7 (35.9 ± 25.5) | 8.0 ± 4.9 | 2.2 ± 0.1 (27.0 ± 8.7) | 77.9 ± 5.2 | 39.4 ± 3.8 (27.0 ± 8.7) | ||
| HUVEC (CCM + Matrigel) | 1:1 (4.7 × 104:1) | 5.2 ± 1.5 | 1.7 ± 0.3 (26 ± 17) | 2.3 ± 0.4 | 0.6 ± 0.3 (25.8 ± 13.1) | 27.1 ± 1.4 | 9.1 ± 0.5 (33.6 ± 2.4) | |
| BT474 | — (13.8 × 104:1) | 0.0 ± 0.9 | — (—) | 0.1 ± 0.1 | — (—) | 2.2 ± 0.7 | — (—) | |
| — (13.8 × 103:1) | 0.2 ± 1.2 | — (—) | 0.0 ± 1.4 | — (—) | 2.2 ± 0.3 | — (—) | ||
| HUVEC (Matrigel) | — (4.7 × 104:1) | 0.5 ± 1.0 | — (—) | 1.0 ± 1.0 | — (—) | 3.3 ± 1.7 | — (—) | |
| HUVEC | — (4.7 × 104:1) | 0.5 ± 1.1 | — (—) | 0.4 ± 1.2 | — (—) | 2.5 ± 0.9 | — (—) | |
| 24 | LnCaP | 1:1 (18 × 104:1) | 17.3 ± 1.8 | 4.5 ± 1.2 (26.2 ± 6.0) | 8.7 ± 1.3 | 2.5 ± 1.0 (29.1 ± 10.9) | 72.6 ± 3.1 | 25.4 ± 1.8 (34.9 ± 2.9) |
| 1:10 (18 × 103:1) | 26.2 ± 5.3 | 7.6 ± 3.8 (29.2 ± 10.2) | 12.4 ± 4.9 | 3.1 ± 2.0 (25.1 ± 12.2) | 73.0 ± 4.1 | 25.6 ± 3.5 (35.01 ± 5.2) | ||
| Mat-Lu | 1:1 (9.6 × 104:1) | 8.4 ± 1.0 | 3.0 ± 0.9 (35.3 ± 12.0) | 7.4 ± 1.3 | 2.5 ± 0.0 (36.0 ± 6.3) | 73.4 ± 2.2 | 24.4 ± 2.7 (33.2 ± 3.7) | |
| 1:10 (9.6 × 103:1) | 11.2 ± 6.0 | 3.8 ± 2.4 (34.3 ± 17.4) | 8.5 ± 4.6 | 2.6 ± 1.3 (30.8 ± 16.8) | 73.9 ± 2.9 | 25.1 ± 1.4 (33.9 ± 2.3) | ||
| HUVEC (CCM + Matrigel) | 1:1 (4.7 × 104:1) | 8.8 ± 2.2 | 2.3 ± 1.3 (23.2 ± 14.2) | 3.2 ± 0.7 | 1.3 ± 0.3 (39.3 ± 12.7) | 23.7 ± 2.5 | 10.9 ± 2.7 (28.6 ± 7.3) | |
| BT474 | — (13.8 × 104:1) | −0.0 ± 0.1 | — | 0.1 ± 0.1 | — | 2.1 ± 0.9 | — | |
| — (13.8 × 103:1) | −0.2 ± 1.6 | — | 0.0 ± 1.4 | — | 2.1 ± 1.3 | — | ||
| HUVEC (Matrigel) | — (4.7 × 104:1) | 0.9 ± 0.9 | — | 0.7 ± 0.9 | — | 2.9 ± 0.6 | — | |
| HUVEC | — (4.7 × 104:1) | 0.4 ± 1.0 | — | 0.7 ± 1.1 | — | 1.9 ± 0.3 | — | |
Specific binding is defined as extent of cell-bound targeted liposomes (or antibody) corrected for extent of nonspecifically bound nontargeted liposomes of same composition (or nonspecifically bound antibody in blocking studies by same nonlabeled antibody). Errors correspond to SD of 2 independent liposome preparations.
CCM = cell-conditioned media.