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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 TOYOPEARL HW-65F base bead via multipoint attachment resulting in excellent base stability for up to 200 CIP cycles with 0.1 mol/L NaOH. 

TOYOPEARL AF-rProtein A HC-650F resin maintains 80% of initial dynamic binding capacity after 40 CIP cycles with 0.5 mol/L NaOH. TOYOPEARL AF-rProtein A HC-650F resin 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.

Achievement of high linear velocities at relatively low pressure enables high throughput at production scale using equipment with moderate pressure limitations. Improved mass transfer characteristics allow it to maintain a larger percent of its capacity at lower residence times relative to agarose base stable resins. Typical leakage for this rProtein A ligand is 0.6 -1.7 ng rProtein A /mg eluted antibody by ELISA testing. Though TOYOPEARL AF-rProtein A-650F resin is shipped in 20% ethanol, it can be stored in 2% benzyl alcohol if necessary.

TOYOPEARL AF-rProtein A HC-650F utilizes an alkaline resistant recombinant Protein A variant. This non-animal derived ligand is bound to the base bead, Toyopearl HW-65F, using multipoint attachment. This contributes to both the resin's alkaline resistance and its low ligand leakages in the range of 5-25ng/mg IgG. Good pressure-flow characteristics and kinetics contribute to maintaining good capacity, particularly at low residence times.

TOYOPEARL AF-rProtein A HC-650F is available in one particle size, 45 µm, which corresponds to the F-grade designation for TOYOPEARL resins.

 

 
P/NDescriptionResin VolumeParticle Size (µm)   Price
($)
 
23425TOYOPEARL AF-rProtein A HC-650F10 mL45  799
23426TOYOPEARL AF-rProtein A HC-650F25 mL45  1,377
23427TOYOPEARL AF-rProtein A HC-650F100 mL45  2,754
23428TOYOPEARL AF-rProtein A HC-650F1 L45  -
23429TOYOPEARL AF-rProtein A HC-650F5 L45  -
23434TOYOPEARL AF-rProtein A HC-650F50 L45  -
       

Product Attributes

Pore size (mean): 100 nm
Particle size (mean): 45 µm
Pressure rating: 0.3 MPa
Shipping buffer: 20% ethanol
pH stability: 2-13
Shelf life (room temp.): 1 year (on going)

Structure:

AFC_rProteinA-HC-650F_structure.png

 
    

Purification of Monoclonal Antibodies


ProA_AF-rProtein-A-HC-650F-Fig8.png

Resin: TOYOPEARL AF-rProtein A HC-650F
Column size: 5 mm ID × 5 cm
Mobile phase: Buffer A: 20 mmol/L sodium phosphate containing 0.15 mol/L NaCl, pH 7.4
Buffer B: 0.1 mol/L citrate, pH 3.0
Flow rate: 61 cm/hr (0.2 mL/min)
Residence time: 5 min
Sample: 40 mL of CHO cell culture, containing 1.0 g/L humanized IgG1



For additional applications, please see our applications database.

 
 
      

Dynamic Binding Capacity

human IgG

ProA_AF-rProtein-A-HC-650F-Fig4.png

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

 

 

 

 


Stability

Base stability

ProA_AF-rProtein-A-HC-650F-Fig2.png

 

Resin: TOYOPEARL AF-rProtein A HC-650F
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 (5 CV)
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

 

 

After CIP with 0.5 mol/L NaOH

ProA_AF-rProtein-A-HC-650F-Fig3.png

 

Resin: TOYOPEARL AF-rProtein A HC-650F
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 (5 CV)
C: 20 mmol/L Na2HPO4, 0.15 mol/L NaCl, pH 7.4 (7 CV)
D: 0.5 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

 

Pressure-Flow Curves

20 cm bed height column

ProA_AF-rProtein-A-HC-650F-Fig5.png

Resin: TOYOPEARL AF-rProtein A HC-650F
Column type: Merck Superformance
Column size: 10 cm ID × 20 cm (1,570 mL)
Mobile phase: DI water
Flow rate: various

 

 

10 cm bed height column

ProA_AF-rProtein-A-HC-650F-Fig6.png

Resin: TOYOPEARL AF-rProtein A HC-650F
Column type: GE Healthcare AxiChrom™
Column size: 20 cm ID × 10 cm (3,140 mL)
Mobile phase: DI water
Flow rate: various

 

Code #DescriptionLiterature Type
AN55 Monoclonal Antibody Purification with a High Capacity Protein A ResinApplication Note
AN61 TOYOPEARL AF-rProtein A HC-650F Host Cell Protein Removal Application Note
AN65 TOYOPEARL AF-rProtein A HC-650F rProtein A Ligand Leakage Application Note
AN73 Purification of a Monoclonal Antibody with TOYOPEARL AF-rProtein A HC-650F at Various Bed HeightsApplication Note
AN78 Comparison of TOYOPEARL AF-rProtein A HC-650F Binding Capacity at Various Bed Heights and Constant Linear VelocityApplication Note
IM02 Instruction Manual for TOYOPEARL and TSKgel PW-type ResinsInstruction Manual
PCAT04 TOYOPEARL and TSKgel Bulk Resin 2016 Product Guide -- Protein AProduct Catalog
PD18 TOYOPEARL AF-rProtein A HC-650F: A High Capacity Protein A Resin Designed to Reduce Production Costs Performance Data
PO38 TOYOPEARL AF-rProtein A HC-650FProduct Overview
RDS3090 TOYOPEARL AF-rProtein A-650F and AF-rProtein A HC-650F ResinsOperating Conditions and Specifications
TP198 Characterization of a Newly Developed High Capacity, Alkaline Resistant, Recombinant Protein A ResinTechnical Presentation
TP203 Comparison of Binding Capacity of TOYOPEARL AF-rProtein A HC-650F Affinity Resin at Varying Bed Heights and Constant Column VolumeTechnical Presentation
TP208 Comparison of Binding Capacity of TOYOPEARL AF-rProtein A HC-650F Affinity Resin at Varying Bed Heights and Constant Column Volume Technical Presentation
P/NDescriptionPrice ($) 
45161MiniChrom TOYOPEARL AF-rProtein A HC-650F859
OC41MDAFPA-650FResin Seeker TOYOPEARL AF-rProtein A HC-650F 96-well plate kit263
23433TOYOPEARL AF-rProtein A HC-650F ELISA919
23430ToyoScreen AF-rProtein A HC-650F, 1 mL690
23431ToyoScreen AF-rProtein A HC-650F, 5 mL (qty. 1)637
23432ToyoScreen AF-rProtein A HC-650F, 5 mL (qty. 5)2,070
45063ToyoScreen RoboColumns AF-rProtein A HC-650F, 200 µL275
45064ToyoScreen RoboColumns AF-rProtein A HC-650F, 600 µL952