Process Separations

Monoclonal Antibody Purification:
Intermediate Purification Resins■■■■■■

Remnant impurities
from capture step
removedClosely related
species resolved
Target mAb
concentration
increased bydecreasing impurities

Ion Exchange Resins for Monoclonal
Antibody Intermediate Purification
Monoclonal antibody (mAb) purification processes
typically involve a multistep workflow consisting of
two or three steps for capture, intermediate, andpolish purification. The resins selected for each ofthese steps must be compatible with the specificpurification challenges that exist at that particularphase of purification.
Intermediate Purification Objectives■■ Remove remnant impurities left over from thecapture step■■

Ideal Features for Intermediate
Purification Resins■■ High binding capacity to get the most efficient
separation of impurities from the mAb■■

Capable of separating/resolving closely related
species and isoforms

Bio-Rad’s Resins for mAb Intermediate
Purification■■ UNOsphere™ Q Anion Exchange Resin■■

Nuvia™ Q Anion Exchange Resin■■

Nuvia™ HR-S Cation Exchange Resin

Increase the target mAb concentration bydecreasing the amount of impuritiesCAPTURE

INTERMEDIATEPOLISH

UNOsphere SUPrA™

UNOsphere Q

CHT™ Ceramic Hydroxyapatite

UNOsphere SUPrA

Nuvia Q

Nuvia™ cPrime™

Nuvia S

Nuvia HR-S

Nuvia cPrime™

UNOsphere Q Anion Exchange Resin
UNOsphere Q is a high-capacity high-throughput anion exchange
resin, based on acrylamido and vinylic monomers. Its largediameter pores and large surface area maximize binding speed,
macromolecule capacity, recovery, and productivity.
Bead Properties
Property

Description

Type of ion exchanger

Strong anion

Functional group–N+(CH3)3Particle size

120 ± 15 μm

Total ionic capacity

120 μeq/ml≥180 mg/ml at 150 cm/hr

Dynamic binding capacity

Recommended linear flow rate

Pressure vs. flow performance≥125 mg/ml at 600 cm/hr
10% BT capacity determined with
2.0 mg/ml BSA in 1.1 x 10 cm column

Competitive Data
Capacity, recovery, and productivity of UNOsphere Q andtwo other anion exchange resins. The pressure/flow propertiesof UNOsphere Q (Bio-Rad Laboratories), Q Sepharose FF(GE Healthcare), and Fractogel EMD TMAE (EMD Millipore)
were evaluated on a 1.1 x 20 cm (20 ml) column equilibrated
with 10 mM Tris buffer, pH 8.5 (buffer A). Bovine serum albumin(BSA; 5.0 mg/ml in buffer A) was loaded until 10% breakthroughoccurred. Elution was performed with buffer A containing 0.5 M
NaCl. Chromatography was performed on a BioLogic DuoFlow™
System. UNOsphere Q exhibits best-in-class DBC, recovery,and productivity, in part due to its open architecture and low
backpressure at high flow rates (Table 1).
Table 1. Comparison of UNOsphere Q, Q Sepharose FF, and Fractogel EMD
TMAE Resin properties.

50–300 cm/hrUnder 2 bar at flow rateof 1,200 cm/hr in DI water(20 x 20 cm packed bed,
1.20 compression factor)

Recovery,%BSA
Binding
Capacity,g/L

Process
Time, hr

Productivity,
g/L/hr

615

100.0

120.0

1.58

75.0

300

99.0

23.0

1.19

19.0

99.0

82.0

5.04

16.0

LinearVelocity,
cm/hr

UNOsphere Q
Q Sepharose FF
Fractogel EMD
TMAE (M)105

SupportCompression factor(settled bed volume/packed bed volume)

1.15 –1.20

pH stability

1–14

Shipping solution

20% ethanol or 0.1 M NaCl

Regeneration

1–2 M NaCl

Sanitization

0.5–1.0 N NaOH

Other Resources■■ Instruction manual, bulletin 4110109

Storage conditions

20% ethanol or 0.1 M NaOH■■

Product information sheet, bulletin 2724
UNOsphere Q Resin technical data, bulletin 2729

Chemical stability
1.0 M NaOH (20°C)

Up to 10,000 hr■■

1.0 M HCI (20°C)

Up to 200 hr■■

Shelf life

5 years

BT, breakthrough; BSA, bovine serum albumin.

Performance Advantages■■ UNOsphere performance — high dynamic binding capacity(DBC) and productivity at high flow rates
■■

■■■■

■■■■

■■

Superior column packing efficiency — high efficiency at very
low flow rates, which remains good at rates up to 1,200 cm/hr;
uniform packing and no channeling or interaction of the sample
with the support
Excellent base stability — little effect on DBC even after
extended storage in both 0.1 N and 1.0 N NaOH
Stable retention times — virtually identical retention times forseveral test proteins at up to 10,000 hr of storage in 1.0 N NaOH
Solvent independent stability — favorable pressure and flow
properties in the presence of common chaotropic agents and at
various pH values and salt concentrations
Optimized bead design — to maximize capture speed ofimpurities from capture eluent or initial feed
Unparalleled quality — for batch-to-batch reproducibility■■

Purification of a monoclonal antibody after Protein A capture,bulletin 5735
Effective cleaning and sanitizing of anion exchange resins,bulletin 5543

Ordering Information
Catalog #Description

Prepacked Screening Tools
732-4714		 Foresight™ UNOsphere Q Plates, 20 μl
732-4819		
Foresight UNOsphere Q RoboColumn Unit, 200 μl732-4820 		
Foresight UNOsphere Q RoboColumn Unit, 600 μl732-4732 		
Foresight UNOsphere Q Column, 1 ml
732-4752 		
Foresight UNOsphere Q Column, 5 ml
Bulk Resin
1560101
1560103156-0105156-0107

UNOsphere Q Support, 25 ml
UNOsphere Q Support, 100 ml
UNOsphere Q Support, 500 ml
UNOsphere Q Support, 10 L

Nuvia Q Anion Exchange Resin
Nuvia Q Resin is an ultra-high capacity high-throughput nextgeneration anion exchange resin. It delivers best-in-classbinding capacity at high flow rates and rapid mass transfer
kinetics, providing a wide experimental design space for processdevelopers.

Competitive Data
DBC and flow rates of Nuvia Q and two other anion exchange
resins. Three 1.1 cm columns were packed to a 10.6 cm bedheight with Nuvia Q, agarose Q, or polymeric Q resins. BSA(5 mg/ml in 20 mM Tris-HCl, pH 8.5) was loaded onto each column
until 10% breakthrough (BT) was observed. Nuvia Q exhibits thehighest DBC of BSA at high linear velocities, relative to the twoother anion exchange resins (Figure 1).

Bead Properties
Description

Type of ion exchanger

Strong anion

Functional group–N+(CH3)3Particle size

85 ± 15 μm

Total ionic capacity100–170 μeq/ml>170 mg/ml at 300 cm/hr

Dynamic binding capacity

10% BT capacity determined with
5 mg/ml BSA in 20 mM Tris-HCl, pH 8.5

Recommended linear flow rate

50–600 cm/hr

Pressure vs. flow performance

Under 3 bar up to 500 cm/hr in DI water

Compression factor(settled bed volume/
packed bed volume)

1.10–1.15

pH stability

Regeneration

1–2 M NaCl

Sanitization

0.5–1.0 N NaOH

Storage conditions

20% ethanol or 0.01 N NaOH

Chemical stability
1.0 N NaOH (20°C)

Up to 1 week

0.01 N NaOH (20°C)

Up to 5 years5 years

BT, breakthrough; BSA, bovine serum albumin.

Performance Advantages■■ Exceptionally high binding capacity — over a range of pHand flow rates

■■

■■

■■

100

0150

Robust stability and performance — stable DBC andrecovery even with extended storage in 1.0 N NaOH; consistent
performance due to chemical stability, ensuring batch-to-batch
reproducibility
Excellent pressure/flow performance — high linear flow rates
without proportional increase in pressure
Improves productivity — contributing to reduced capital costs,
space requirements, and cycle time
Unsurpassed quality — for batch-to-batch reproducibility
Flexible format — usable in both affinity-based and affinityindependent workflows300

450600Linear velocity, cm/hr

Fig. 1. Comparison of DBC vs. flow velocity of Nuvia Q Resin and two other anion
exchange resins. Nuvia Q Resin ( u ); agarose Q resin ( n ); polymeric Q resin ( s ).

Other Resources■■ Instruction manual, bulletin 10018215
■■

■■

■■

150

50

Long term: 4–12
20% ethanol + 0.1 M NaCl

■■

200

Short term: 2–14

Shipping solution

Shelf life

250

DBC BSA, 10% BT, mg/mlProperty

Product information sheet, bulletin 6129
Automated mAb workflows: combining multidimensional(Multi-D) purifications with product analysis, bulletin 6745

Ordering Information
Catalog #Description

Prepacked Screening Tools
732-4703 		
Foresight Nuvia Q Plates, 2 x 96-well, 20 μl
732-4804 		
Foresight Nuvia Q RoboColumn Unit, 200 μl
732-4805 		
Foresight Nuvia Q RoboColumn Unit, 600 μl
732-4721 		
Foresight Nuvia Q Column, 1 x 1 ml
732-4741 		
Foresight Nuvia Q Column, 1 x 5 ml
Bulk Resin
15604111560413156-0415
156-0417

Nuvia Q Media, 25 mlNuvia Q Media, 100 ml
Nuvia Q Media, 500 ml
Nuvia Q Media, 10 L

Nuvia HR-S Cation Exchange Resin
Nuvia HR-S Resin is a small particle high-resolution cationexchange resin designed for intermediate and final polishpurification steps where the separation of closely related
biomolecules is challenging. Its exceptional selectivity and highrecovery is due to optimized particle size and chemistry. It is
designed to meet biomolecule purification needs at both thelaboratory- and large bioprocess-scale levels.

Bead Properties
Property

Description

Type of ion exchanger

Strong cationFunctional group–SO3–Particle size

50 ± 10 μm

Total ionic capacity

100–180 μeq/ml

cation exchanger, Resin 1. Nuvia HR-S delivered a final aggregate
content of <0.3% and a recovery of >80% while the recovery using
agarose-based Resin 1 was <70% for the same final aggregate
content (Figure 2). The higher capacities and better recovery
translate to reduced cost of goods for downstream processes.

≥70 mg/ml at 300 cm/hr

Recommended linear flow rate

Pressure vs. flow performance

Compression factor(settled bed volume/packed bed volume)

10% BT capacity determinedwith 5.0 mg/ml hIgG in 20 mM
Na acetate, pH 5.0.
50–200 cm/hr
Under 2 bar at flow rateof 200 cm/hr in DI water

1.6Aggregate, %Dynamic binding capacity

Short term: 2–14

0.8
0.4

(20 x 20 cm packed bed,
1.25 compression factor)

00

1.20–1.25

pH stability

1.2102030

40

50

60

7080

Fig. 2. Performance of Nuvia HR-S ( ) vs. Resin 1 ( ).

Long term: 4–13

Shipping solution

20% ethanol

Regeneration

1–2 M NaClSanitization

0.5–1.0 N NaOH

Storage conditions

20% ethanol or 0.1 N NaOH

Chemical stability

Other Resources■■ Instruction manual, bulletin 10033316■■■■

1.0 N NaOH (20°C)

Up to 5 weeks (840 hr)

0.1 N NaOH (20°C)

Up to 5 years

Shelf life

5 years

Product information sheet, bulletin 6448
Improving aggregate removal from a mAb feedstream using
high-resolution cation exchange chromatography, bulletin 6439

Ordering Information

BT, breakthrough; hIgG, human immunoglobulin G.

Catalog #Performance Advantages■■ High binding capacity — over a wide operating pH range

Prepacked Screening Tools
732-4707 		
Foresight Nuvia HR-S Plate, 20 μl
732-4831 		Foresight Nuvia HR-S RoboColumn Unit, 200 μl
732-4832 		
Foresight Nuvia HR-S RoboColumn Unit, 600 μl
732-4723 		
Foresight Nuvia HR-S Column, 1 ml
732-4743 		
Foresight Nuvia HR-S Column, 5 ml

■■

■■

■■

■■

90

Monomer recovery, %Excellent pressure/flow performance — linear relationshipbetween pressure and flow up to 200 cm/hr
Superior resolution — able to separate mAb aggregatesfrom monomers
Consistent and reproducible performance — no decrease inBSA DBC and recovery when exposed to 840 hr of 1.0 N NaOH;
consistent performance due to chemical stability, ensuringbatch-to-batch reproducibility and repetitive use withoutcompromising performance
Excellent scalability — compatibility from bench tomanufacturing steps

Competitive Data
Aggregate content and monomer recovery. Aggregate
clearance and monomer recovery of 46 mg/ml of IgG/ml resin wascarried out using Nuvia HR-S and an agarose-based small particle

Bulk Resin
15605111560513156-0515
156-0517

Description

Nuvia HR-S Media, 25 ml
Nuvia HR-S Media, 100 ml
Nuvia HR-S Media, 500 ml
Nuvia HR-S Media, 10 LAll our resins come with full regulatory support backed by
Bio-Rad’s global application and development team. Contact
your regional Bio-Rad process chromatography specialistat process@bio-rad.com or call customer service at
1-800-4-BIORAD (1-800-424-6723) for more information.
Test drive our resins for your mAb purification.
Visit bio-rad.com/web/ResinSample to order your sample.
Fractogel is a trademark of Merck KGaA. Sepharose is a trademark ofGE Healthcare. RoboColumn is a trademark of Atoll GmbH.

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