100 4 100

Flo 4 Flo
w (1 w (1
00, 150 5 00, 150 5
300 ) 300 .9)
) 200 (4.9 ) 200 6 . (4
6 d. nd
250 Co
n
250 7 Co
7
300 300 8
8
130
10 9 9
130
10

150
30

10% DBC
150

10% DBC
30

10% DBC
170
50
170
50
190
70
190

CHROMATOGRAPHY
4 70 4
5 100 5 100
6 150 6 150
.9) 7 200 .9) 7 200 Flo
. (4 Flo . (4

Nuvia™ cPrime™ Hydrophobic

w (1 nd w (1
nd 8 8 00,
Co 250
300
00, Co 250
300
300 9 ) 300 9 )

Cation Exchange Resin

■■ New selectivity New Selectivity and Large Design Space for Downstream
Salt tolerance
Purification Processes
■■

■■ Simple method
development
■■  arge design space for
L Introduction
binding and elution Nuvia cPrime Hydrophobic Cation Exchange Resin is a new addition to Bio-Rad’s expanding family of
■■ High recovery mixed-mode purification products. Nuvia cPrime provides unique selectivity, high recovery, and versatility
■■ Mechanical and in large-scale purification applications for a variety of therapeutic proteins. Nuvia cPrime is effective for
chemical stability initial capture and polish applications, especially for molecules that present a purification challenge using
current schemes.

Mixed-Mode Ligand
Nuvia cPrime Resin is designed with a mixed-
mode ligand (Figure 1) that provides a unique H
balance between hydrophobic and charged N
characteristics. The ligand structure also provides O
an opportunity for hydrogen-bonding interactions. H
N
Importantly, the balance of weak acid and OH
hydrophobic components is optimized to allow
for straightforward method development and O
predictable behavior during binding and elution. Fig. 1. Mixed-mode ligand for Nuvia cPrime Resin.

Properties of Nuvia cPrime Resin

Property Description
Functional group Hydrophobic weak cation exchange
Base matrix composition Macroporous highly crosslinked polymer
Particle size 70 µm ± 10 μm
Dynamic binding capacity* (hlgG) >40 mg/ml
Dynamic binding capacity (lactoferrin) >60 mg/ml
Ligand density 110–150 μeq/ml
Recommended linear flow rate 50–600 cm/hr
Pressure vs. flow performance** Under 2 bar at a flow rate of 600 cm/hr
pH stability
Short-term 3–14
Long-term 4–13
Chemical stability 1.0 N NaOH, 8 M urea, 6 M GuHCl, 6 M KSCN, 3 M NaCl,
1% Triton X-100, 2% SDS + 0.25 M NaCl, 20% EtOH,
70% EtOH, 30% IPA
Shipping solution 20% ethanol
Storage conditions 20% ethanol
Shelf life*** 5 years
* At 10% breakthrough, 300 cm/hr.
** 20 × 20 cm packed bed (1.17 compression factor).
*** Stored at room temperature in 20% ethanol.
 New and Unique Selectivity Chromatographic Performance
The balance between hydrophobicity, charged for Novel Therapeutics
interaction, and the highly hydrophilic base matrix Nuvia cPrime is effective for the purification of established
of Nuvia cPrime empowers method developers therapeutic proteins as well as the increasingly diverse
with new ways to directly exploit various modes new constructs that are in development (many of which
of interaction to purify challenging or sensitive lack an affinity handle). Salt- and pH-sensitive proteins
proteins or to separate closely related protein with a high propensity for aggregation and/or degradation
species, such as isoforms and variants from post- can now be effectively purified using simplified methods.
translational modifications, product aggregation,
and degradation (Figure 2). Lysozyme binding capacity, mg/ml

0) Binding pH (4, 8)
, 40
2.00 — Conductivity
Elution of
300
(10
target protein M
m
— A 280 Regenerate l],
1.75
250 aC
[N 50
1.50 ing

Conductivity, mS/cm
nd
Flowthrough/impurities 200
Bi

cit y, mg /ml
1.25 45
1.00 150
40
AU

0.75
100

Binding capa
35
0.50
50 30

ml
0.25

pacity, mg/
0.00 0 25

0.00 60.00 120.00

Binding ca

) 00
Min, tenth

(10, 4
Chromatographic

l], mM
30
Condition Specification 20
25

g [NaC
Column 0.56 x 4 cm
Flow rate 300 cm/hr
Loading buffer 50 mM sodium phosphate + 150 mM NaCl (pH 6.5)

Bindin
Elution buffer 50 mM sodium phosphate + 400 mM NaCl (pH 7.0) Binding
pH (4,
Equilibration Loading buffer 8)
Wash Loading buffer
Lysozyme recovery, %
Elution Elution buffer
Elution pH (4, 8)
Regenerate 1 N NaOH 0)
0
,0
,1
Fig. 2. Chromatographic performance and unique selectivity. (10 0
M
Nuvia cPrime Resin enables effective separation. AU, absorbance units. l],
m
aC
[N
n
io
Large Design Space for Binding and Elution ut 30
El

Recover y, %
Nuvia cPrime is designed for versatile capture and high
recovery across a wide range of salt concentrations
and pH (Figure 3). These properties may allow for direct 40

loading without the need for dilution. Integrating

a Nuvia cPrime step into a multicolumn process
%

is operationally simple. 60
ver y,

)
0
Reco

1,0 0
(10,
mM

80
Cl],
[Na

100
ion

Elution
Elut

pH (4, 8
)

Fig. 3. Large design space afforded by Nuvia cPrime Resin.

© 2014 Bio-Rad Laboratories, Inc. Bulletin 6242

Simple Method Development Built to Meet the Demands
The mixed-mode nature of the Nuvia cPrime ligand and of Commercial Operations
its associated range of interactions allows for a directed Nuvia cPrime is built on a porous polymeric base matrix
and intuitive approach to method development and that delivers low backpressure at high flow rates (Figure 5).
process optimization. Alternatively, a simple design of It is also chemically and mechanically stable. Fast mass
experiment (DOE) exercise will quickly guide developers transfer dynamics ensure efficient chromatography at
to optimum loading, wash, and elution conditions high flow, making Nuvia cPrime Resin an operationally
afforded by the resin’s large design space (Figures superior choice for commercial-scale applications.
3 and 4).

1 2 3
Elution by pH increase Elution by varying conductivity* Elution using modifiers

■■ Consider mobile
phase modifiers
[Salt]

Elution not achieved Elution not achieved

■■ Consider using a
different salt
■■ Consider a different pH
within the range
[Salt]
pH

Elution achieved Elution achieved

■■  efine method using

R ■■  efine method using salt
R ■■  efine method using
R
pH gradient gradient at a given pH modifier or different salt
Elution achieved ■■ Convert gradient into ■■ Convert gradient into ■■ Convert gradient into
step elution protocol step elution protocol step elution protocol

* At optimum pH, determined from step 1.

Fig. 4. Recommended approach to method development.

A. Column backpressure vs. linear velocity B. Dynamic binding capacity vs. linear velocity, lactoferrin
3.0 80

2.5 70
10% BT DBC, mg/ml

60
2.0
Pressure, bar

50
1.5
40
1.0
30
0.5
20
0.0 10
–0.5 0
0 200 400 600 800 1,000 1,200 150 300 450 600
Linear velocity, cm/hr Linear velocity, cm/hr

Fig. 5. Nuvia cPrime displays low backpressure at high flow rates. A, flow performance of Nuvia cPrime Resin in a Bio-Rad ® InPlace™
Column. A 20 x 20 cm column with 17% axial compression was used. B, dynamic binding capacity vs. linear velocity of Nuvia cPrime Resin.
A 1.1 x 9.6 cm column was loaded with 5.25 mg/ml lactoferrin in 20 mM NaOAc + 150 mM NaCl, pH 4.5, until 10% breakthrough was
observed. BT, breakthrough; DBC, dynamic binding capacity.

© 2014 Bio-Rad Laboratories, Inc. Bulletin 6242

 Applications and Regulatory Support
Regulatory support files and application notes are available
upon request. Bio-Rad Laboratories is an ISO 9001 registered
corporation.
Please visit us at bio-rad.com/nuvia for more information about
Bio-Rad’s complete line of process chromatography solutions.
Ordering Information
Catalog # Description
156-3401 Nuvia cPrime Hydrophobic Cation Exchange Resin, 25 ml
156-3402 Nuvia cPrime Hydrophobic Cation Exchange Resin, 100 ml
156-3403 Nuvia cPrime Hydrophobic Cation Exchange Resin, 500 ml
156-3404 Nuvia cPrime Hydrophobic Cation Exchange Resin, 1 L
156-3405 Nuvia cPrime Hydrophobic Cation Exchange Resin, 5 L
156-3406 Nuvia cPrime Hydrophobic Cation Exchange Resin, 10 L
732-4705 Foresight™ Nuvia™ cPrime™ Plates, 20 μl
732-4807 Foresight Nuvia cPrime RoboColumn Unit, 200 μl
732-4808 Foresight Nuvia cPrime RoboColumn Unit, 600 μl
732-4722 Foresight Nuvia cPrime Column, 1 ml
732-4742 Foresight Nuvia cPrime Column, 5 ml

Larger volumes are available on request.

Triton is a trademark of Dow Chemical Company.

Bio-Rad
Laboratories, Inc.

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Bulletin 6242 Rev C US/EG 14-2336 1214 Sig 1214