02

Ion Exchange Columns
and Media

BioPro IEX Columns----------------------------------- 24-26
BioPro IEX SmartSep Q/S and BioPro IEX Q/S --------- 27-29
Ordering Information------------------------------------- 30

02

Ion Exchange Columns and MediaIon exchange columns

BioPro IEX Columns
Usable pH range : 2-12

Ion Exchange Columns and Media ‌Hydrophilic polymer beads with low nonspecific adsorption ‌Ultra-fast analysis and high resolution analysis on non-porous type ‌Ideal for analysis and laboratory scale purification on porous type with high binding capacity
and high recovery of biomolecules
 S‌ uitable for characterization of biopharmaceuticals and quality control

Ion exchange columns for separation of proteins, peptides, and nucleic acids
BioPro IEX columns are specially designed for separation of proteins, peptides, and nucleic acids.
BioPro IEX QF/SF are based on non-porous hydrophilic polymer beads with high chemical and mechanical stability, and low
nonspecific adsorption of biomolecules. The short columns (30 mm, 50 mm) are useful for the fast analysis at a higher flow rate, andthe 100 mm length columns are best choice for the quality control assessment of biopharmaceuticals requiring a high resolution.
BioPro IEX QA/SP are based on porous hydrophilic polymer beads. BioPro IEX QA/SP have superior resolution, high binding
capacity and high recovery of various biomolecules, and they allow highly effective analysis and laboratory scale purification ofbiopharmaceutical proteins such as antibodies.

SEM images of polymer beads

Non-porous polymer beads

Porous polymer beads

SpecificationsBioPro IEX QF
Matrix3−CH2N（CH3）Counter ion
Ion exchange capacity*(meq/mL-resin)245−CH2CH2CH2SO3+−CH2N（CH3）
3−CH2CH2CH2SO3−

Cl−

Na+

Cl−

Na+0.09

0.24

0.09

0.09

>12（BSA）>10（human-IgG）>110（BSA）>70（human-IgG）−

Usable temperature

4-60℃

Usable pH range

2-12

Column material

PEEK*Reference value

BioPro IEX SP

Hydrophilic porous polymer

3, 5+Dynamic binding capacity*(mg/mL-resin)

BioPro IEX QA

Hydrophilic non-porous polymer

Particle size (μm)
Charged group

BioPro IEX SF

High resolution analysis of monoclonal antibody (MAb) <1>
BioPro IEX SF

3 µm, 100 X 4.6 mmI.D.

Eluent: A) 20 mM NaH2PO4-Na2HPO4 (pH 6.8)
B) 20 mM NaH2PO4-Na2HPO4 (pH 6.8)
containing 0.2 M NaCl
0-50%B (0-30 min)Flow rate: 0.5 mL/min (180 cm/hr)
Temperature : 25°C
Detection: UV at 215 nm
Injection: 10 μL
Sample: Humanized monoclonal IgG1 (2.5 mg/mL)

1200
1000
800
600
400200

BioPro IEX SF column with 3 µm, 100 mm length achieved
high resolution analysis of MAb.

U170221C30510152025min

Ion Exchange Columns and MediamAU

High resolution analysis of monoclonal antibody (MAb) <2>
BioPro IEX SF

5 µm, 100 X 4.6 mmI.D.

tR 60.0 min

mAU
12mAU
128tR 60.0 min

84404004020Competitor’s
WCX column
mAU
mAU

3mAU
321282112mAUEluent

2160

1030
4003080
min406080

V110221F00 V110221F 20mAU3mAU
3250607080 min

50607080 minmin

10 µm, 250 X 4.0 mmI.D.

tR 118.2 min

The separation of MAb is compared on BioPro IEX SF andcompetitor’s column under the same gradient conditions atpH 5.6. BioPro IEX SF column provides higher resolution ofMAb in a shorter analysis time than the competitor's column.

tR 118.2 min

8410904V110223G00900 V110223G 2000: A) 20 mM MES-NaOH (pH 5.6)
B) 20 mM MES-NaOH (pH 5.6)
containing 0.2 M NaCl
Initial gradient conc. : 35%B (70 mM NaCl)
Gradient slope: 0.25%B/min (0.5 mM NaCl)
Flow rate: 180 cm/hr(0.5 mL/min for 100 X 4.6 mmI.D.,
0.378 mL/min for 250 X 4.0 mmI.D.)
Temperature: 30°CDetection: UV at 280 nm
Injection: 10 μL
Sample: Humanized monoclonal IgG1 (1 mg/mL)

20110

130min110
4060

130min80100120140 min

4060

80

100120140 min

Excellent batch-to-batch reproducibility

ColumnEluentBatch E
Batch E
Batch D
Batch D
Batch C
Batch C
Batch B
Batch B

: BioPro IEX SF 5 μm, 100 X 4.6 mmI.D.
: A) 20 mM NaH2PO4-Na2HPO4 (pH 6.5)
B) 20 mM NaH2PO4-Na2HPO4 (pH 6.5)
containing 0.2 M NaCl
0-50%B (0.5-30 min)Flow rate: 0.5 mL/min (180 cm/hr)
Temperature : 25°CDetection: UV at 215 nm
Injection: 10 μL
Sample: Humanized monoclonal IgG1

BioPro IEX SF column exhibits excellent batch-to-batch
reproducibility for MAb analysis, including the resolution of small
peaks for charge variants. All the medium batches are inspectedby various quality control tests and must pass rigorous criteria
before release. BioPro IEX columns are the best choice for thequality control of MAbs and other biopharmaceuticals.

Batch A051015202530Batch A min

051015202530min

25

02

Ion Exchange Columns and MediaHigh resolution analysis of nucleic acids
BioPro IEX QF

5 µm, 100 X 4.6 mmI.D.

DNA fragments 1Kb DNA ladder (75-12,216 bp)

Ion Exchange Columns and MediaEluent: A) 20 mM Tris-HCl (pH 8.1) containing 0.7 M NaClB) 20 mM Tris-HCl (pH 8.1) containing 1.0 M NaCl
0-100%B (0-30 min)Flow rate: 0.5 mL/min (180 cm/hr)
Temperature : 25°C
Detection: UV at 260 nm
Injection: 20 μL (0.25 mg/mL)

mAU

50

0N080212G

01020 minThe separation of DNA fragments is shown. BioPro IEX QF of 100 mm
length column is a good choice for high-resolution analysis of nucleicacids.

High binding capacity and recovery
Comparison of dynamic binding capacity (DBC) and recovery for BSA
DBC（mg/mL-resin, 10% breakthrough）

Eluted amount（mg/mL-resin）

Recovery*（%）

BioPro IEX QA

126

120

95

Brand T (porous Q type)

73

58

79

Brand G (porous Q type)1003535*Recovery:（Eluted amount/DBC）X 100

Breakthrough curves

0.25Column: BioPro IEX QA, 50 X 4.6 mmI.D.
Brand T (porous Q type) 50 X 4.6 mmI.D.
Brand G (porous Q type) 50 X 5.0 mmI.D.
Linear velocity: 180 cm/hr
Equilibration buffer : 20 mM Tris-HCl (pH 8.6)
Elution buffer: 20 mM Tris-HCl (pH 8.6) containing 1.0 M NaClDetection: UV at 280 nm
Sample: 1 mg/mL Bovine serum albumin (BSA) inequilibration buffer

BioPro IEX QA

Brand GBrand TAU0.50

10% breakthrough0020406080100120140

BSA amount loaded (mg/mL-resin)

160

BioPro IEX QA gives the superior DBC and recovery compared with conventional porous polymer anion exchange columns. The surface structure of porous
type BioPro IEX, which is designed for maximum interaction with proteins, provides high binding capacity, and the hydrophilic property of polymer beads
significantly reduces nonspecific adsorption of proteins.

High loadability
Comparison of the effect of sample load on BioPro IEX QA and commercial Q type column
BioPro IEX QA5 μm, 50 X 4.6 mmI.D.

Brand G (porous Q type)10 μm, 50 X 5.0 mmI.D.

1. Ovalbumin
2. Trypsin inhibitor

1mAU

21Loadingamount200
N080206B

100

250 μgN080207C

50 μg

N080206A

0102030 min

2100

250 μgN080208C

100 μgN080208B

050 μgN080206D

03×1072×107
1×107010: A) 20 mM Tris-HCl (pH 8.1)
B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl
10-80%B (0-30 min)Flow rate: 0.5 mL/min (180 cm/hr for 4.6 mmI.D., 150 cm/hr for 5.0 mmI.D.)
Temperature : 25°CDetection: UV at 280 nm
Injection: 100 μL

20

BioPro IEX QA
Brand G(porous Q type)

4×107
500 μgN080206E

100 μgN080207B

0200

500 μgRecovery of trypsin inhibitor

5×107

Peak areamAU

30 min

00.5

1.0

1.5

Loading (mg)2.0Eluent

26

BioPro IEX QA shows excellent resolution and peak shapes evenwhen the loading amount increases. The porous type BioPro IEX
columns are suitable for laboratory-scale purification of proteins.

2.5

Ion exchange media

Usable pH range : 2-12

 ‌High productivity on purification ‌Hydrophilic polymer beads with low nonspecific adsorption ‌High binding capacity and high resolution over a wide range of flow rate ‌Suitable for purification of antibodies, proteins and nucleic acids

Ion exchange media for purification of biopharmaceuticals
BioPro IEX SmartSep Q/S and BioPro IEX Q/S, strong ion exchange media, are suitable for downstream chromatographic purification
processes in biopharmaceutical manufacturing. High dynamic binding capacity and high recovery of those media allow fast purification
processes at large scale. It offers high productivity on industrial purification.
BioPro IEX SmartSep Q/S, which show high resolution and recovery even at a high flow rate and high loading condition, are suitable
for an intermediate purification step and a polishing step of MAbs, proteins, peptides, and oligonucleotides.
BioPro IEX Q/S are designed for capture and intermediate purification of biopharmaceuticals such as MAbs.

Specifications

Ion Exchange Columns and Media

BioPro IEX SmartSep Q/S
BioPro IEX Q/SCounter ion

BioPro IEX SmartSep Q

BioPro IEX SmartSep S

MatrixBioPro IEX Q

BioPro IEX S

Hydrophilic porous polymer

Particle size (μm)
Charged group10, 20, 30+−R-N（CH3）3Ion exchange capacity(meq/mL-resin)
Dynamic binding capacity(mg/mL-resin)

75+−R-N（CH3）
3−R-SO3−>0.08>100（BSA）−R-SO3−>0.10>160（BSA）>100（lysozyme）

Usable pH range>160（lysozyme）

2-12

DBC (mg/mL-resin)High dynamic binding capacity (DBC) for various samples

120

BioPro IEX SmartSep S30100

Brand G（porous S type 30 μm）

Conditions of DBC measurement*Column: 50 X 5.0 mmI.D.
Flow rate: 400 cm/hr (1.32 mL/min)
Temperature : 25°C

Brand T（porous S type 30 μm）

80*Inquire us for details.

60
40
200Insulin(MW 5,800)

Lysozyme(MW 14,300)

Human Polyclonal IgG(MW 150,000)

DBC（mg/mL-resin, 10% breakthrough）

Particlesize（μm）

InsulinLysozyme

Human Polyclonal IgG

BioPro IEX SmartSep S30

307311193

Brand T（porous S type）

30677242

Brand G（porous S type）

30648541

BioPro IEX media have higher DBC compared to conventional ion exchange media. Especially for IgG, BioPro IEX media have more than twice as high DBC
as competitors' media. This feature of BioPro IEX media makes purification productivity of IgG per unit time double or more.

27

02

Ion Exchange Columns and MediaDBC over a wide range of flow rate
BioPro IEX SmartSep S30
Brand T (porous S type 30 µm)

110Brand G (porous S type 30 µm)

10090

DBC (mg/mL-resin)

Ion Exchange Columns and Media

1208070Column: 50 X 5.0 mmI.D.
Equilibration buffer : 20 mM citric acid-NaOH (pH 5.3)
Elution buffer: 0.5 M NaCl in equilibration buffer
Flow rate: 200-800 cm/hr (0.66-2.62 mL/min)
Temperature: ambient (25℃)
Detection: UV at 280 nm
Sample: 1.5 mg/mL human polyclonal IgGin equilibration bufferHigh DBC of BioPro IEX media is maintained even at higher flow
rate, making them suitable for the high-speed purification with 2-3
times of conventional flow rates. This feature offers significant
improvement on productivity.

60504030
20100200400600

800

1000

Linear velocity (cm/hr)

Excellent durability (Stability on CIP)
Test protocols
Separation of standard proteins
Determination of DBC and recovery
CIP with 1 M NaOH(5 column volumes)

DBC and recovery
120200

DBC

100
Recovery

150

80

10060

50

2Conditions of DBC* measurement
Column: BioPro IEX S75, 50 X 5.0 mmI.D.
Flow rate: 800 cm/hr (2.62 mL/min)
Equilibration buffer : 20 mM Glycine-NaOH (pH 9.0)
Elution buffer: 0.5 M NaCl in equilibration buffer
Sample: 1.0 mg/mL Lysozyme in equilibration buffer
Temperature: ambient
Detection: UV at 300 nm

468101214161820

Recovery (%)

DBC (mg/mL-resin)

250*DBC was determined at 10% breakthrough40

Number of CIP with 1 M NaOH

Separation of standard proteins
32

1

Number of CIP

20th
10th

1. Ribonuclease A
2. Cytochrome c
3. Lysozyme

Conditions of separation of standard proteins
Column: BioPro IEX S75, 50 X 5.0 mmI.D.
Eluent: A) 20 mM NaH2PO4-Na2HPO4 (pH 6.8)
B) 20
‌ mM NaH2PO4-Na2HPO4 (pH 6.8)
containing 0.5 M NaClGradient: 0-100%B (0-60 min; Linear)
Flow rate: 180 cm/hr (0.59 mL/min)
Temperature : 25°CDetection: UV at 220 nmInjection: 24 µL (0.5 mg/mL)

initial
1020304050 min

Cleaning in place (CIP) is an important procedure for cleaning and sterilization of columns used for protein purification. The DBC and the selectivity of proteins
are unaffected following 20 cycles of CIP with 1 M NaOH. The high chemical stability of BioPro IEX media allow effective cleaning with alkaline solution.

28

Excellent batch-to-batch reproducibility of DBC
250

DBC (mg/mL-resin)

200
150100
BioPro IEX S75

500

BioPro IEX Q75

Batch A

Batch B

Batch C

Batch DBatch E: 50 X 4.6 mml.D.
: 180 cm/hr

for anion-exchange resin
Equilibration buffer : 20 mM Tris-HCI (pH 8.6)
Elution buffer: 0.5 M NaCl in equilibration buffer
Sample: 1.5 mg/mL BSA in equilibration buffer
Detection: UV at 280 nm
for cation-exchange resin
Equilibration buffer : 20 mM Glycine-NaOH (pH 9.0)
Elution buffer: 0.5 M NaCl in equilibration buffer
Sample: 1.5 mg/mL Lysozyme in equilibration bufferDetection: UV at 300 nm

BioPro IEX media exhibit excellent batch-to-batch reproducibility of DBC. All the medium batches are inspected by various quality control tests. We supply
stable products over a long period of time.

Ion Exchange Columns and Media

ColumnFlow rate

BioPro Ion Exchange Screening Kit
The BioPro Ion Exchange Screening Kit is a set of screening
columns packed with BioPro IEX media.
・Two column types (1 mL and 5 mL) ideal for media screening,development of purification methods, and loadability studies
・Easy installation and convenient use

29

02

Ion Exchange Columns and Media
Ordering Information –Columns–
BioPro IEX QF/SF
Column length (mm)Ion Exchange Columns and Media

Phase
dimension

Column I.D.
(mm)

30

50

100

BioPro IEX QF
non-porous
3 μm

4.6

QF00S03-0346WP

QF00S03-0546WPQF00S03-1046WP

BioPro IEX QF
non-porous
5 μm

4.6

QF00S05-0346WP

QF00S05-0546WPQF00S05-1046WP

BioPro IEX SF
non-porous
3 μm

4.6

SF00S03-0346WP

SF00S03-0546WP

SF00S03-1046WP

BioPro IEX SF
non-porous
5 μm

4.6

SF00S05-0346WP

SF00S05-0546WP

SF00S05-1046WP

Phasedimension

Column I.D.
(mm)

3050100

BioPro IEX QAporous
5 μm

4.6

QAA0S05-0346WPQAA0S05-0546WPQAA0S05-1046WP

BioPro IEX SP
porous
5 μm

4.6

SPA0S05-0346WPSPA0S05-0546WPSPA0S05-1046WP

BioPro IEX QA/SPColumn length (mm)Ordering Information
BioPro IEX media
Product name

Particle size (μm)

BioPro IEX SmartSep Q10

Product number
QSA0S1010BioPro IEX SmartSep S10
BioPro IEX SmartSep Q20

SSA0S10QSA0S2020BioPro IEX SmartSep S20
BioPro IEX SmartSep Q30

SSA0S20QSA0S3030

BioPro IEX SmartSep S30
BioPro IEX Q75

SSA0S30QAA0S75

75

BioPro IEX S75

SPA0S75

BioPro Ion Exchange Screening Kit
Packing material

Particle size (μm)

Specification

BioPro IEX SmartSep Q20205/pack

BioPro IEX SmartSep S20
BioPro IEX SmartSep Q30305/pack

BioPro IEX SmartSep S30
BioPro IEX Q7575
BioPro IEX S75305/packColumn volume (mL)

Product number

1

BPQSA0S20-01PK

5

BPQSA0S20-05PK

1

BPSSA0S20-01PK

5

BPSSA0S20-05PK

1BPQSA0S30-01PK

5BPQSA0S30-05PK

1

BPSSA0S30-01PK

5

BPSSA0S30-05PK

1

BPQAA0S75-01PK

5

BPQAA0S75-05PK

1

BPSPA0S75-01PK

5

BPSPA0S75-05PK