SOLUTIONS

Protein A for antibody purification

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.

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
Protein A structure

Product attributes

Pore size (mean): 100 nm
Particle size (mean): 45 µm (F-grade)
Pressure rating: 0.3 Mpa
Shipping buffer: 20% ethanol
pH stability: 2-13
Shelf life (room temperature): 1 year (ongoing)

DBC of of TOYOPEARL AF-rProtein A HC-650F
Dynamic binding capcity of TOYOPEARL Protein A resin

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
Comparison of residence time and capacity of TOYOPEARL Protein A resins

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 
Base stability of TOYOPEARL Protein A resin 
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 

 

Ordering Information

P/N Description Dimension
0023425 TOYOPEARL AF-rProtein A HC-650F 10 mL
0023426 TOYOPEARL AF-rProtein A HC-650F 25 mL
0023427 TOYOPEARL AF-rProtein A HC-650F 100 mL
0023428 TOYOPEARL AF-rProtein A HC-650F 1 L
0023429 TOYOPEARL AF-rProtein A HC-650F 5 L
0045063 RoboColumn AF-rProtein A HC-650F 200 µL x 8 
0045064 RoboColumn AF-rProtein A HC-650F 600 µL x 8
0045161 MiniChrom AF-rProtein A HC-650M, 5 mL 8 mm ID x 10 cm L
0023430 ToyoScreen AF-rProtein A HC-650F 1 mL x 5
0023431 ToyoScreen AF-rProtein A HC-650F 5 mL x 1
0023432 ToyoScreen AF-rProtein A HC-650F 5 mL x 5
0022899 Protein A-R40 Standard  10 mg/L (0.5 mL) 

Corresponding ELISA kit is supplied by Cygnus Technologies:
https://cygnustechnologies.com/product_detail/tosoh-r40-and-r28-protein-a-mix-n-go-elisa.html