VirCap® InertShell Core-Shell Resin
For Adenoviridae ADV Purification
VirCap® InertShell is designed with core-shell technology where the beads are designed and optimized for the purification of viruses and other large biomolecules. The core-shell technology allows for dual functionality combining size-exclusion separation with binding chromatography. Viruses and other large biomolecules that are too large to penetrate the inert shell of the chromatography resins are collected in column flow-through fraction (FT mode). Contaminants (< Mr 700 000), on the other hand, pass through the inert outer shell and bind to the ligands in the inner core.
VirCap® InertShell is made of polymethacrylate microspheres with octylamine ligand, as Pic 1. The shell of the microspheres is neutral and hydrophilic which has nonspecific protein adsorption. The pore size of the shell (50-100 nm) is smaller than that of the core (200-500 nm). And the thickness of the shell is about 0.5-1.0μm. The shell prevents proteins (molecular weights greater than 700 kDa) from entering the core. In the chromatography process, large-size inactivated viruses cannot enter the microsphere core that they are breakthrough and be collected quickly. The octylamine ligand in the core will realize the dual functions of anion exchange and hydrophobicity that it can quickly capture protein molecules with molecular weight less than 700 kDa. VirCap® InertShell can effectively remove cell protein of the host, DNA fragment, endotoxin, albumin, and other impurities in the pseudo-purified system.
Parameter of VirCap® InertShell
|VirCap® Inert Shell||Competitor 700|
|Matrix||Polyacrylate||Highly cross‑linked agarose|
|Average particle size||50-150 μm||50-150 μm|
|Density of ligand||0.10-0.20 mmol/mL||0.04-0.085 mmol/mL|
|Binding capacity¹||20 mg BSA/mL resin||12 mg BSA/mL resin|
|Operational pressure||≤1.0 MPa||≤0.3 MPa|
|Operational flow rate||100-600 cm/h||100-600 cm/h|
|Chemical stability||All commonly used aqueous buffers, 1 M sodium hydroxide (NaOH)², 6 M guanidine hydrochloride, 30% isopropanol, and 70% ethanol.|
|Storage||20% ethanol at 4°C to 25 ℃|
- Dynamic binding capacity measured at 5% breakthrough with 76 cm/h on φ10×13 mm, 1 mL columns. The buffer was 1.0 mg/mL BSA 50 mM NaCl, pH 0.
- No significant changes in ionic capacity and carbon content after storage 1 week in 1 M NaOH at 25°C.
Compared with Competitor 700, the thickness of the core in VirCap® Inert Shell (0.5-1.0 μm) is smaller than Competitor 700 (5 μm). VirCap® Inert Shell is conducive to the rapid mass transfer of impure proteins to the medium core for capture, like cell proteins of the host, DNA fragments, endotoxin, serum. VirCap® Inert Shell has a higher yield amount of miscellaneous protein. With the macroporous structure (200-500 nm) of the core in VirCap® Inert Shell can quickly remove the miscellaneous protein in CIP. VirCap® Inert Shell also has a longer service life in the purification process. In animal vaccine studies, the performance of VirCap® Inert Shell can be repeated use more than 30 times with less change in its properties.