Bulk packings
MacroSep IEX Q are innovative strong anion exchange media to target large biomolecules and particles such as adeno-associated viruses (AAVs), protein complexes, and plasmid DNA (pDNA). This media offers an optimized macro-porous structure that ensures efficient purification of these molecules. With its high binding capacity for large particles and molecules in addition to high separation efficiency even at elevated flow rates, MacroSep IEX Q is an ideal solution for downstream processing (DSP) platforms, enabling successful and productive purification of large biotherapeutics.
MacroSep IEX Q
BioPro IEX SmartSep Q30
| Specifications | |
| Matrix | Hydrophilic methacrylate polymer |
|---|---|
| Particle size (µm) | 30 |
| Pore size (Å) | 9000 |
| Charged group | -R-N+(CH3)3 |
| Ion-exchange capacity (meq/mL-resin) |
>0.08 |
| Dynamic binding capacity (DBC) (mg/mL-resin) |
30 (Thyroglobulin) |
| Usable pH range | 2-12 |
The direct comparison of MacroSep IEX Q with conventional media at two different flow rates clearly shows that MacroSep IEX Q provides excellent resolution, even at high flow rates. This leads to high productivity.
| Column | 50 X 5.0 mmI.D. |
|---|---|
| Eluent | A) 20 mM Bis-tris propane-HCl (pH 9.0) B) 20 mM Bis-tris propane-HCl containing 0.5 M TMAC* (pH 9.0) |
| Gradient | 10-50%B (45 CV) |
| Temperature | 25°C |
| Detection | FLS at Ex. 280 nm, Em. 348 nm |
| Injection | 30 µL |
| Sample | AAV2 (1.07 X 1011 vg/mL) |
* tetramethylammonium chloride
This research was supported by AMED under Grant Number JP18ae0201001.
AAV Full/Empty capsids separation was performed with MacroSep IEX Q. Purity of the full AAV2 after the chromatographic purification was reached to 91.2% against 54.4% of a feed solution, determined by analytical ultracentrifugation (AUC). MacroSep IEX Q is suitable for chromatographic purification of AAV.
| Column | MacroSep IEX Q (30 µm), 50 X 5.0 mmI.D. |
|---|---|
| Eluent | A) 20 mM Bis-tris propane-HCl (pH 9.0), 1% sucrose, 0.1% poloxamer 188 B) 20 mM Bis-tris propane-HCl containing 0.5 M CC* (pH 9.0), 1% sucrose, 0.1% poloxamer 188 |
| Gradient | 0-60%B (36 CV) |
| Flow rate | 1.0 mL/min |
| Temperature | ambient (25°C) |
| Detection | UV at 280 nm |
| Injection | 48 mL |
| Sample | AAV2 (0.72 X 1011 vg/mL) |
* choline chloride
This research was supported by AMED under Grant Number JP18ae0201001.
MacroSep IEX Q can separate supercoiled (SC) from the non-functional linearized (L) and nicked open-circular (OC) species with higher resolution while conventional media gives poor resolution of those species. MacroSep IEX offers increased productivity with higher purity on pDNA purification.
| Column | 50 X 5.0 mmI.D. |
|---|---|
| Eluent | A) 20 mM Tris-HCl (pH 8.5) B) 20 mM Tris-HCl (pH 8.5) containing 1.0 M NaCl |
| Gradient | [MacroSep IEX Q] 82%B (0-1.5 min), 82-86%B (1.5-22.5 min) [Brand B] 77%B (0-1.5 min), 77-81%B (1.5-22.5 min) |
| Flow rate | 0.4 mL/min |
| Temperature | 25°C |
| Detection | UV at 260 nm |
| Injection | 10 µL (60 µg/mL) |
| Sample | [SC] pUC19 plasmid extracted from E. coli (2686 bp) [OC] pUC19 plasmid digested with nicking endonuclease Nt.BspQΙ [L] pUC19 plasmid digested with restriction enzyme BamHΙ |
MacroSep IEX Q is designed to have higher DBC value of large biomolecules than conventional media for improving productivity. DBC value of pDNA with each MacroSep IEX Q and conventional media is determined as an example of a large-sized modality. MacroSep IEX Q shows higher DBC with high recovery, compared to the conventional media.
| Column | MacroSep IEX Q, 26 X 7 mmI.D. (1 mL)
Brand C, 20 X 8 mmI.D. (1 mL) |
|---|---|
| Equilibration buffer | 100 mM CH3COONa-CH3COOH (pH 5.0) |
| Elution buffer | Equilibration buffer containing 1 M NaCl |
| Flow rate | 0.5 mL/min (Residence time 2 min) |
| Temperature | ambient |
| Detection | UV at 260 nm |
| Sample | pRP[Exp]-Puro-TRE3G>EGFP (4934 bp) |
| Sample Concentration | 0.2 mg/ml pDNA in Equilibration buffer containing 0.15 M NaCl |
| DBC (mg/mL-resin, 10% breakthrough) |
Recovery* (%) |
|
|---|---|---|
MacroSep IEX Q |
7.5 | 101 |
Brand C |
1.3 | 101 |
*Recovery: (Eluted amount/Adsorption amount) X 100
