TOYOPEARL AF-rProtein A HC-650F
TOYOPEARL AF-rProtein A HC-650F is a high capacity Protein A resin for monoclonal antibody purification. An enhanced rProtein A ligand is bound to the polymethacrylate base bead via multipoint attachment resulting in excellent base stability for up to 200 CIP cycles with 0.1 mol/L NaOH. It maintains 80% of its initial dynamic binding capacity after 40 CIP cycles with 0.5 mol/L NaOH. TOYOPEARL AF-rProtein A HC-650F also tolerates exposure to above ambient temperatures.
TOYOPEARL AF-rProtein A HC-650F is available in 45 µm particle size (F-grade) and exhibits dynamic binding capacities of greater than 65 g/L at 5 minutes residence times and greater than 50 g/L at 2 minutes residence time with feed stock concentrations from 1.0 g/L to 10.0 g/L. Improved mass transfer characteristics allow it to maintain a larger percent of its capacity at lower residence times relative to comparable agarose based resins. It is therefore ideally suited for continuous operation using multiple short columns. This resin is also available in pre-packed SkillPak columns to simplify your process developmentand scale up.
TOYOPEARL AF-rProtein A HC-650F utilizes an alkaline resistant recombinant Protein A variant. This new non-animal derived ligand is bound to the base bead using multipoint attachment. This contributes to both the new resin's alkaline resistance and its low ligand leakages in the range of 5-25 ng ligand/mg IgG.
| Structure of TOYOPEARL AF-rProtein A HC-650F |
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Product attributes
| Pore size (mean): |
100 nm |
| Particle size (mean): |
45 µm (F-grade) |
| Shipping buffer: |
20% ethanol |
| pH stability: |
2-13 |
| Stability data: |
1 year (ongoing) |
| DBC of of TOYOPEARL AF-rProtein A HC-650F |
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Resin: TOYOPEARL AF-rProtein A HC-650F
Column size: 5 mm ID × 5 cm
Mobile phase: 0.02 mol/L sodium phosphate, 0.15 mol/L NaCl, pH 7.4
Residence time: 2, 3.5, 5 min
Detection: UV @ 280 nm (10% breakthrough)
Sample: human IgG @ 1, 5, 10 g/L in mobile phase |
| Comparison of residence time and capacity |
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Resins: TOYOPEARL AF-rProtein A HC-650F
TOYOPEARL AF-rProtein A-650F
MabSelect SuRe™ LX
Column size: 5 mm ID ×5 cm
Mobile phase: 0.02 mol/L sodium phosphate, 0.15 mol/L NaCl, pH 7.4
Residence time*: 2, 3.5, 5 min
Detection: UV @ 280 nm
Sample: human IgG @ 1 g/L in mobile phase
*MabSelect SuRe DBC data taken from product brochure (2, 4, and 6 minutes residence times).
DBC was calculated at 10% breakthrough |
| Base stability of TOYOPEARL AF-rProtein A HC-650F |
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Column size: 5 mm ID × 5 cm
Wash procedure: A: 20 mmol/L Na2HPO4, 0.15 mol/L NaCl, pH 7.4 (10 CV)
B: 0.1 mol/L citrate, pH 3.0 (5CV)
C: 20 mmol/L Na2HPO4, 0.15 mol/L NaCl, pH 7.4 (7 CV)
D: 0.1 mol/L NaOH (3 CV – 15 min contact time)
E: 20 mmol/L Na2HPO4, 0.15 mol/L NaCl, pH 7.4 (5 CV)
Capacity: DBC was determined at 10% breakthrough after every 20 cycles |