Table 1.
List of Materials Used during Downstream Process Development of Lentivirus Vector Production
| Process Step | Support | Material/MWCO | Manufacturer | Notes |
|---|---|---|---|---|
| Clarification/filtration | Millistak+ Pod depth filter CE50, 0.027 m2, ref. MCE50027H1 | cellulose | Merck Millipore | standard in small-scale runs |
| Sartopore 2 midcap size 7: 0.8 + 0.45 μm, 0.05 m2, ref. 5445306G7 | polyethersulfone (PES) | Sartorius Stedim Biotech | tested | |
| Sartoclean CA midcap size 7: 0.45 μm, 0.08 m2, ref. 5625306A7 | cellulose | Sartorius Stedim Biotech | tested | |
| Millistak+ Pod depth filter CE50, 0.77 m2, ref. MCE5007FS1 | cellulose | Merck Millipore | first and second scale-up run (single filter) | |
| TFF concentration and buffer exchange | Sartocon Slice Hydrosart 0.11 m2, 100 kDa, ref. 305 144 6801E-SG | cellulose/hydrosart | Sartorius Stedim Biotech | used in TFF1a development and in the scale-up runs TFF2b (single cassette) |
| Sartocon Hydrosart 0.6 m2, 100 kDa, ref. 302 144 6806E-SG | cellulose/hydrosart | Sartorius Stedim Biotech | 5 × 0.6 m2 in first scale-up run TFF1 | |
| Sartocube Hydrosart 3 m2, 100 kDa, ref. 302 144 6830E-BSW | cellulose/hydrosart | Sartorius Stedim Biotech | 1 Sartocube + 2 × Sartocon (total 4.2 m2) in second scale-up run TFF1 | |
| Pellicon 2 Mini Ultracel type V 0.1 m2, 100 kDa, ref. P2C100V01 | cellulose/Ultracel | Merck Millipore | tested for TFF1 | |
| Pellicon 2 Mini Ultracel type V 0.1 m2, 300 kDa, ref. P2C300V01 | cellulose/Ultracel | Merck Millipore | tested for TFF1 | |
| Pellicon 2 Mini Biomax type V 0.1 m2, 100 kDa, ref. P2B100V01 | PES/Biomax | Merck Millipore | tested for TFF1 | |
| Chromatography | CIMmultus QA 1 mL (2 μm), ref. 311.5113-2 | CIM monolithic advanced composite column | BIASeparations | tested for AEXc |
| CIMmultus QA 1 mL (6 μm), ref. 311.5113-6 | CIM monolithic advanced composite column | BIASeparations | tested for AEX | |
| CIMmultus DEAE 1 mL (2 μm), ref. 311.5114-2 | CIM monolithic advanced composite column | BIASeparations | tested for AEX | |
| Sartobind Q nano 3 mL, 8 mm, ref. 96IEXQ42EUC11-A | reinforced cellulose | Sartorius Stedim Biotech | standard in small-scale runs | |
| Sartobind Phenyl nano 3 mL, 8 mm, ref. 96HICP42EUC11-A | reinforced cellulose | Sartorius Stedim Biotech | tested for HIC | |
| Mustang Q XT Acrodisc 0.86 mL, ref. MSTGXT25Q16 | hydrophilic PES | Pall | tested for AEX | |
| Sartobind Q 150 mL, 8 mm, ref. 96IEXQ42E9BFF | reinforced cellulose | Sartorius Stedim Biotech | AEX/first scale-up run | |
| Sartobind Q 400 mL, 8 mm, ref. 96IEXQ42E1HSS | reinforced cellulose | Sartorius Stedim Biotech | AEX/second scale-up run |
The list includes clarification, tangential flow filtration, and chromatographic supports. DEAE, diethylaminoethyl; HIC, hydrophobic intercation chromatography; MWCO, molecular weight cutoff.
a
Concentration and buffer change after clarification using tangential flow filtration.
b
Final buffer change using tangential flow filtration.
c
Anion exchange chromatography.