Table 2.
Degradation Half-Lives of Oligonucleotides in Serum
| Hours (95% confidence interval) | ||||||
|---|---|---|---|---|---|---|
| Identifier | Frozen human | Fresh human | Fresh mouse | Preincubated frozen human | Preincubated fresh human | Preincubated fresh mouse |
| DNA T | 16 (12–23) | 8.2 (7.2–9.4) | 1.8 (1.6–1.9) | |||
| DNA C | 6 (4.8–7.6) | 4.9 (4.4–5.4) | 1.6 (1.4–1.9) | |||
| fYrR T | 9.9 (6.8–15) | 12 (9.3–15) | 2.5 (2.1–2.8) | 53 (34–110) | 56 (34–13) | 9.9 (6.7–14) |
| fYrR C | 11 (9.9–13) | 10 (8.4–13) | 2.2 (2.0–2.4) | 53 (40–74) | 59 (52–68) | 3.7 (3.3–4.3) |
| fGmH T | >240 | >240 | 180 (150–250) | |||
| fGmH C | >240 | >240 | 110 (85–150) | |||
| OMe T | >240 | >240 | 230 (130–830) | |||
| OMe C | >240 | >240 | 75 (60–99) | |||
| A9.min | 8.0 (6.1–11) | 3.8 (2.7–5.3) | 2.5 (2.4–2.7) | |||
The degradation half-life of A9.min and C36 constructs in frozen human, fresh human, and fresh mouse serum presented in h, including the 95% confidence interval. Preincubated serum was heated to 37°C for 5 days before addition of oligonucleotides. The fraction of intact oligonucleotide was determined from a plot of the area corresponding to the full-length oligonucleotide on the gel. Half-life calculations are the result of a minimum of two independent experiments. The potency of fresh and frozen human serum begins to decrease after 24 h; hence half-life calculations are based on DyLight 650 measurements taken in the first 24 h assuming 100% degradation over time. Mouse serum retains its potency longer; half-lives in mouse serum are based on DyLight [650 measurements taken over 72 h (fresh mouse serum)] or 48 h (preincubated mouse serum).