Process Catalog 2014 - page 61

57 Tosoh bioscience PROCESS
AFC
Ultra-high capacity
TOYOPEARL AF-rProtein A HC-650F
TOYOPEARL AF-rProtein A HC-650F is the newest affinity
resin introduced by Tosoh Bioscience. It exhibits dynamic
binding capacities of greater than 65 g/L at residence times
of 5 minutes 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
(Figure 2).
Improved mass transfer characteristics allow it to maintain
a larger percent of its capacity at lower residence times
(Figure 3) relative to agarose based, caustic stable resins.
The multipoint attachment of the enhanced recombinant
protein A ligand to the TOYOPEARL matrix is resulting in
excellent base stability for up to 200 CIP cycles with 0.1
mol/L NaOH at 15 min contact time (Figure 4). It maintains
80% of initial dynamic binding capacity after 40 CIP cycles
with 0.5 mol/L NaOH (Figure 5).
Protein a Affinity
chromatography
figure 2
Column: TOYOPEARL AF-rProtein A HC-650F (5 mm ID × 5 cm L)
Mobile phase: 20 mmol/L sodium phosphate, 150 mmol/L NaCl pH 7.4;
Residence time: 2, 3.3, 5 min; Detection: UV @ 280 nm
Sample: polyclonal human IgG @ 1, 5, 10 g/L in mobile phase
DBC measured at 10 % breakthrough
DBC at various loads and residence times
figure 3
Column: TOYOPEARL AF-rProtein A HC-650F (5 mm ID x 5 cm L)
Mobile phase: 20 mmol/L sodium phosphate, 150 mmol/L NaCl pH 7.4;
Residence time: 2, 3.3, 5 min; Detection: UV @ 280 nm;
Sample: polyclonal human IgG @ 1 g/L in mobile phase;
DBC measured at 10 % breakthrough. MabSelect SuReTM and MabSelect
SuReTM LX DBC data from GE brochure. MabSelect SuReTM and MabSelect
SuReTM LX are registered trademarks of GE Healthcare Bio-Sciences AB,
Uppsala, Sweden.
DBC of high capacity Protein A media
figure 4
Column size: 5 mm ID × 5 cm L; Wash procedure: A: 20 mmol/L Na2 HPO4
0.15 mol/L NaCl, pH 7.4 (10 CV)
B: 0.1 mol/L citrate, pH 3.0 (5 CV)
C: 20 mmol/L Na2HPO 4, 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 Na2 HPO4, 0.15 mol/L NaCl, pH 7.4 (5 CV)
CIP Study with 0.1 M NaOH