Table 2.
Comparison of the Harvesting Methods.
| Steps | Lysis A | TrypLE | Lysis B | Lysis C | Lysis D |
|---|---|---|---|---|---|
| Collected supernatant | Y | Y | Y | Y | Y |
| Chemical lysis buffer with benzonase treatment (1 h) | Y | N | N | N | N |
| Washes (1–2 x) | N | Y | Y | Y | Y |
| TrypLE | N | Y | N | N | N |
| Mechanical disassociation (firmly tapped 5–10 x) | N | Y | Y | Y | Y |
| TrypLE | N | Y | N | N | N |
| Centrifugation | N | 1000 rpm | 1000 rpm | 1000 rpm | 1000 rpm |
| Supernatant removed & discarded | N | Y | Y | Y | Y |
| Pellet saved & resuspended in 8–10 mL | N | Y | Y | Y | Y |
|
Chemical lysis buffer with benzonase
(1 h) on pellet |
N | N | Y | Y | N |
| Pipette-based disruption and homogenization | N | Y | Y | Vortexed | Vortexed |
| Frozen −80 °C, thawed 3 x | N | Y | Y | Y | Y |
| Centrifugation | 3000 rpm | 4000 rpm | 4000 rpm | 4000 rpm | 4000 rpm |
| Supernatant retained, 5% sucrose, & frozen at −80 °C | Y | Y | Y | Y | Y |
| Viral titer by ICC | Y | Y | Y | Y | Y |