AbSolute® is based on a modified silica matrix that exhibits strong mechanical, thermal and chemical stability. The matrix is rigid and incompressible. These exceptional mechanical properties combined with unique kinetic performances allow operations at higher velocities (up to 1000 cm/h) and the use of higher bed lengths, hence using smaller diameter columns and reducing capital investment and cleanroom space requirements.
The particles are small (44 μm) with a perfect spherical shape and narrow size distribution. Average pore size is 1000 Å and shows narrow size distribution (+/- 150 Å).
Optimized particles and pores allow high efficiency and faster mass transfer which increases the Dynamic Binding Capacity (DBC).
To read our AbSolute® , silica-based protein A, Scientific Poster: click here!
Breakthrough curves at 200, 300 and 600 cm/h were performed with industrial mAb broth to determine dynamic binding capacities at 10% breakthrough of Media X and AbSolute®.
Media were packed in columns of 5 mm I.D. and 50 mm bed height; the bed volume was 0.98 mL. The concentration of IgG in industrial broth was 1.5 g/L. During the loading, fractions were collected and analyzed by HPLC to determine IgG content.
Breakthrough with AbSolute® occurs later than with media X and the breakthrough curve for AbSolute® is very steep; this is the result of uniform particles and homogenous pore size.
Fig1: Breakthrough curves
AbSolute® provides much higher Dynamic Binding Capacity (DBC) at 10% breakthrough than media X.
At 200, 300 and 600 cm/h, the Dynamic Binding Capacities (DBC) of AbSolute® were respectively 38%, 76% and 100% higher than the DBC of media X. Very high dynamic capacity is reached at high linear flow rates and low-residence time.
Fig2: DBC at 10% Breakthrough of Media X and Absolute®
Fig3: DBC versus Residence Time
*Obtained with polyclonal IgG
N.B.: Media X = 3rd generation agarose – based Protein A media.
AbSolute® is incompressible, leading to a linear relationship between back pressure and velocity.
The combination of rigid structure and uniform particle size allows operation at velocities up to 1000 cm/h with a bed length higher than 20 cm.
Fig4: Pressure Drop versus Linear Velocity
IgG purified by AbSolute® has similar purity and impurity profile than the same IgG purified by media X (demonstrated with various IgGs and mAbs).
AbSolute® is easy to pack and can be used with most chromatography equipment (low, medium and high pressure) from lab to production scale.
For instance, the packing of a 20 cm bed length in a 110 mm I.D. Prochrom®-Bio column takes 2 minutes to obtain 5900 plates/m at 160 cm/h.
Using AbSolute® allows greater flexibility in column selection, bed length optimization and process design. With great experience in silica packing, Novasep guarantees both the scale-up and the process performance.
The change of DBC during 150 cycles including CIP 50 mM NaOH was evaluated.
The testing conditions were:
– Cleaning in place: 50 mM NaOH
– Residence time = 20 min
– Cleaning volume = 5, 10 and 20×C.V. during 150 cycles.
– Column: 4.6×10 mm (0.17 mLbed)
– Loading: 0.2 mg/mL polyclonal human IgG in PBS (pH=7.4), 20 min; Washing: PBS, 10 min; Elution: 0.15 M citric acid containing 0.15 M NaCl (pH=2.2), 10 min
After 150 cycles including cleaning in place with 50 mM NaOH, the DBC is 87% of the initial DBC
Fig5: Change of DBC during 50 mM NaOH Cleaning
|Support||Spherical Silica, modified for high pH resistance|
|Average Particle Size||44 μm|
|Specific Surface Area||65 m2/g|
|Pore Diameter||1000 Å|
|Ligand||Recombinant, Protein A - free of animal derived products|
|Static Binding Capacity||70 mg polyclonal human IgG/mL-Bed|
|Dynamic Binding Capacity||≥ 40 mg/mL DBC at 10% breakthrough at 250 cm/h with 5x100 mm I.D.xL|
column (residence time 2.4 minutes)
≥ 29 mg/mL DBC at 10% breakthrough at 600 cm/h with 5x50 mm I.D.xL
column (residence time 0.5 minutes)
≥ 20 mg/mL DBC at 10% breakthrough at 1000 cm/h with 5x50 mm I.D.xLDBC
column (residence time 0.3 minutes)
|Working pH (except CIP)||1.5 - 9.0|
|Recommended CIP||100 mM NaOH + 0.5 M NaCl every 10 cycles
6M Urea every 10 cycles
|Delivery Condition||0.1 M sodium acetate buffer containing 1% benzyl alcohol pH 5.2|
|up to 1000 cm/h|
|Recommended Bed Height||up to 40 cm (depending on application and equipment pressure rating)|
|Storage Temperature||+2°C - 6°C /36°F - 43°F (do not freeze)|
|Bioburden||≤ 10 CFU/mL|
|Endotoxin||≤ 2 EU/mL|
AbSolute® is manufactured and quality controlled in a cGMP compliant manufacturing facility. AbSolute® is available in several volume sizes.
AbSolute® must be stored at +2°C to +6°C (36°F - 43°F).
For more details, please download the AbSolute® brochure.
Range of biochromatography solutions:
Range of TFF solutions:
Download our brochures for biologics manufacturing.
For any questions on Novasep's purification/separation services and equipment for biologics, please contact us.