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.

NOW AVAILABLE - NEW PROTEIN A ELISA KIT optimized for TOYOPEARL AF-rProtein A HC-650F and TOYOPEARL AF-rProtein A-650F resins
Tosoh R40 and R28 Protein A, Mix-N-Go ELISA Kit from Cygnus Technologies, catalog # F910. (https://cygnustechnologies.com/product_detail/tosoh-r40-and-r28-protein-a-mix-n-go-elisa.html)

P/N	Description	Resin Volume	Particle Size (µm)	Price ($)
0023425	TOYOPEARL AF-rProtein A HC-650F	10 mL	45	693	Inquire
0023426	TOYOPEARL AF-rProtein A HC-650F	25 mL	45	1,194	Inquire
0023427	TOYOPEARL AF-rProtein A HC-650F	100 mL	45	2,387	Inquire
0023428	TOYOPEARL AF-rProtein A HC-650F	1 L	45	-	Inquire
0023429	TOYOPEARL AF-rProtein A HC-650F	5 L	45	-	Inquire

Product Attributes

Pore size (mean):	100 nm
Particle size (mean):	45 µm
Shipping buffer:	20% ethanol
pH stability:	2-13
Stability data (room temp.):	5 years

Structure:

Purification of Monoclonal Antibodies

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

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

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

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

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

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 #	Description	Literature Type
AN055	Monoclonal Antibody Purification with a High Capacity Protein A Resin	Application Note
AN061	TOYOPEARL AF-rProtein A HC-650F Host Cell Protein Removal	Application Note
AN065	TOYOPEARL AF-rProtein A HC-650F rProtein A Ligand Leakage	Application Note
AN073	Purification of a Monoclonal Antibody with TOYOPEARL AF-rProtein A HC-650F at Various Bed Heights	Application Note
AN078	Comparison of TOYOPEARL AF-rProtein A HC-650F Binding Capacity at Various Bed Heights and Constant Linear Velocity	Application Note
AN116	Reduce Downstream Processing Costs for mAbs by Switching to a Tosoh 2-Step Platform	Application Note
BR10	Best-In-Class Bioprocessing Resins	Brochure
IM02	Instruction Manual for TOYOPEARL and TSKgel PW-type Resins	Instruction Manual
PCAT04	TOYOPEARL, TSKgel and Ca⁺⁺Pure-HA Bulk Resin 2019 Product Guide -- Affinity	Product 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-650F	Product Overview
RIS0009	TOYOPEARL AF-Protein-A-HC-650F	Resin Information Sheet
TP198	Characterization of a Newly Developed High Capacity, Alkaline Resistant, Recombinant Protein A Resin	Technical Presentation
TP203	Comparison of Binding Capacity of TOYOPEARL AF-rProtein A HC-650F Affinity Resin at Varying Bed Heights and Constant Column Volume	Technical 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
TP226	Protein A Chromatography: Important Features in Purification to Reduce mAb Aggregation and Increase Recovery	Technical Presentation
TP229	Two-Step Purification Process for Mouse Monoclonal Antibody from Hybridoma Cell Line	Technical Presentation

P/N	Description	Price ($)
OC41MDAFPA-650F	Resin Seeker TOYOPEARL AF-rProtein A HC-650F 96-well plate kit	-	Inquire
0045228	SkillPak Antibody 1 mL column library	434	Inquire
0045232	SkillPak Best in Class 1 mL column library	430	Inquire
0045266	SkillPak Best in Class 5 mL column library	1,406	Inquire
0045229	SkillPak mAb Platform 1 mL column library	430	Inquire
0045263	SkillPak mAb Platform 5 mL column library	844	Inquire
0045201	SkillPak TOYOPEARL AF-rProtein A HC-650F 1 mL column	133	Inquire
0045222	SkillPak TOYOPEARL AF-rProtein A HC-650F 1 mL columns (qty. 5)	662	Inquire
0045258	SkillPak TOYOPEARL AF-rProtein A HC-650F 5 mL column	384	Inquire
0023430	ToyoScreen AF-rProtein A HC-650F, 1 mL (qty. 5)	-	Inquire
0023431	ToyoScreen AF-rProtein A HC-650F, 5 mL (qty. 1)	-	Inquire
0023432	ToyoScreen AF-rProtein A HC-650F, 5 mL (qty. 5)	-	Inquire
0045063	ToyoScreen RoboColumns AF-rProtein A HC-650F, 200 µL	-	Inquire
0045064	ToyoScreen RoboColumns AF-rProtein A HC-650F, 600 µL	1,418	Inquire