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. 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
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
0045228 SkillPak Antibody 1 mL col. Library
(AF-rProtein A HC-650F, AF-rProtein L-650F, NH2-750F,
Sulfate-650F, GigaCap Q-650M, GigaCap S-650S 1 mL x 1 ea)
7 mm ID x 2.5 cm L
0045229 SkillPak mAb Platform 1 mL col. library
(AF-rProtein A HC-650F, Sulfate-650F, NH2-750F 1 mL x 2 ea)
7 mm ID x 2.5 cm L
0045263 SkillPak mAb Platform 5 mL col. library
(AF-rProtein A HC-650F, Sulfate-650F, NH2-750F 5 mL x 1 ea)
8 mm ID x 10 cm L
0045232 SkillPak Best in Class 1 mL col. Library
(AF-rProtein A HC-650F, AF-rProtein L-650F, NH2-750F,
Sulfate-650F, Ca++Pure-HA 1 mL x 1 ea)
7 mm ID x 2.5 cm L
0045266 SkillPak Best in Class 5 mL col. library
(AF-rProtein A HC-650F, AF-rProtein L-650F, NH2-750F,
Sulfate-650F, Ca++Pure-HA 5mL x 1 ea)
8 mm ID x 10 cm L
0045201 SkillPak 1 AF-rProtein A HC-650F 1 mL col. 7 mm ID x 2.5 cm L
0045222 SkillPak 1 AF-rProtein A HC-650F 5x1 mL col. 7 mm ID x 2.5 cm L
0045258 SkillPak 5 AF-rProtein A HC-650F 5 mL col. 8 mm ID x 10 cm L
0045338 SkillPak 50 AF-rProtein A HC-650F 50 mL col. 2.5 cm x 10 cm
0045339 SkillPak 200 AF-rProtein A HC-650F 200 mL col. 5 cm x 10 cm
0045063 RoboColumn AF-rProtein A HC-650F 200 µL x 8
0045064 RoboColumn AF-rProtein A HC-650F 600 µL x 8 
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