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PRODUCT BULLETinPOROS® XS Cation Exchange Resin
POROS® XSCation Exchange ResinHigh capacity + high resolution + high salt tolerance
• Dynamic binding capacity for proteins, greater than 100 mg/mL at 5% breakthrough
• High resolution using optimized 50 μm particle size for improved impurity clearance
• Robust salt tolerance that maintains dynamic binding capacity up to150 mM naCl (15 mS/cm)
4.50 5.00 5.50 6.00 6.50
75.00
112.50
150.00
120110
100
90
80
70
60
40
30
20 0
80
90
100
10
50
37.50
0.00
0
20
40
60
80
100
120
140
160
0 20 40 60 80 100 120 140 160
Salt concentration (mM)
Conductivity (mS/cm)
10 12 14 1686420
pH 4.5 pH 5.0
100 cm/H300 cm/H700 cm/H
0
1000
2000
3000
4000
5000
6000
mAU
2500
2000
1500
1000
500
30.0 40.0 50.0 60.0 70.0 ml
0
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
500
1000
1500
2000
2500
401
POROS® HS POROS® XS
Volume (mL)
Volume (mL) Volume (mL)Volume (mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Abso
rban
ce a
t 280
nm
(mAU
)6000
4000
5000
3000
2000
1000
50 10 15 20 25 30 35 40
0
Abso
rban
ce a
t 280
nm
(mAU
)
Capa
city
at 5
% b
reak
thro
ugh
(mg/
mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Salt Concentration (mM)
pH
• Design Points
Salt
Conc
entr
atio
n (m
M)
Figure 1. 5% Breakthrough IgG binding capacity vs. salt concentration and pH.High capacity is obtained under a wide range of process conditions. The area inside the dotted line indicates the condition at which greater than 100 mg/mL binding capacity of IgG could be obtained. Column: 0.46 cmD x 20 cmL; Buffer: 20 mM MES; Load conditions: 5 mg/mL IgG; Flow rate: 300 cm/h
Introduction
Today’s higher-titer biotherapeutic processes create a host of downstream purification challenges and bottlenecks. Traditionally, high-capacity resins fail to achieve high resolution in the purification of monoclonal antibodies and recombinant proteins. in addition, maintaining protein binding capacity in the presence of higher salt concentrations is a hurdle in streamlining process flow.
The need for a resin that delivers high capacity and high resolution in a wide range of salt concentrations is now critical, especially in downstream polishing steps.
A resin that delivers the combination of high capacity, high resolution, and high salt tolerance can lead to increased process efficiency and help enable a faster time to market for the product.
Figure 2. Separation capability on POROS® HS and POROS® XS resin. Column: 1 cmD x 20 cmL; Gradient: 10-50% B, 7.5 CV; Buffer A: 20 mM MES, 25 mM NaCl pH 6.2; Buffer B: 20 mM MES, 1M NaCl pH 6.2; Flow rate: 300 cm/hSample: Chymotrypsinogen, Cytochrome C, Lysozyme
Figure 3. Resolution over a range of operating flow rates. Column: 0.46 cmD x 20 cmL; Gradient: 10–100% B, 16 CV; Buffer A: 20 mM MES, 25 mM NaCl pH 6.2; Buffer B: 20 mM Tris, 1.0 M NaCl, pH 6.2; Flow rate: 300 cm/hSample: Chymotrypsinogen, Cytochrome C, Lysozyme
POROS® XS Cation Exchange Resin
POROS® XS resin, from Life Technologies, is the first high-capacity, high-resolution resin that allows loading in the presence of up to 15 mS/cm naCl to more than 100 mg/mL capacity, while delivering unprecedented separation capability.
A major improvement on traditional cation exchangers, POROS® XS resin sets the standard for cation exchange chromatography.
• Dynamic binding capacity leads to reduced column size, smaller footprint, decreased buffer usage, and reduced cycling.
• High resolution supports better product yield and improved impurity clearance.
• Robust salt tolerance allows improved process flexibility, and alignment and process flow streamlining.
• Low back pressure and linear pressure response help improve scalability.
• Rigid polymeric bead with covalent surface chemistry makes for easier handling and packing, and superior physical and chemical stability, resulting in a robust manufacturing process.
Dynamic binding capacity
POROS® XS resin delivers high capacity, greater than 100 mg/mL at 5% breakthrough, over a broad range of process conditions. Figure 1 demonstrates unparalleled high binding capacity across a broad range of pH and salt conditions. Higher dynamic binding capacity was achieved through the optimization of the base bead pore structure and the surface chemistry.
High resolution
With an optimized 50 μm particle size, POROS® XS resin maintains the superior resolution capability of POROS® HS resin (Figure 2) while addressing the challenging impurity levels associated with high titer processes. Resolution is consistent across a wide range of operating flow rates (Figure 3) and maintained independent of protein load (Figure 4).
Robust salt tolerance
POROS® XS resin delivers high and consistent protein capacity across a broad range of salt concentrations (Figure 5) up to 150 mM naCl (15 mS/cm conductivity). The flexibility of loading columns, under higher conductivity conditions, helps enable the reduction or elimination of the dilution of column feed streams or the removal of unit operations, such as tangential flow filtration. The optimized surface ionic capacity of POROS® XS resin minimizes competitive binding between the protein of interest and the counterion, resulting in superior salt tolerance.
4.50 5.00 5.50 6.00 6.50
75.00
112.50
150.00
120110
100
90
80
70
60
40
30
20 0
80
90
100
10
50
37.50
0.00
0
20
40
60
80
100
120
140
160
0 20 40 60 80 100 120 140 160
Salt concentration (mM)
Conductivity (mS/cm)
10 12 14 1686420
pH 4.5 pH 5.0
100 cm/H300 cm/H700 cm/H
0
1000
2000
3000
4000
5000
6000
mAU
2500
2000
1500
1000
500
30.0 40.0 50.0 60.0 70.0 ml
0
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
500
1000
1500
2000
2500
401
POROS® HS POROS® XS
Volume (mL)
Volume (mL) Volume (mL)Volume (mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Abso
rban
ce a
t 280
nm
(mAU
)
6000
4000
5000
3000
2000
1000
50 10 15 20 25 30 35 40
0
Abso
rban
ce a
t 280
nm
(mAU
)
Capa
city
at 5
% b
reak
thro
ugh
(mg/
mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Salt Concentration (mM)
pH
• Design Points
Salt
Conc
entr
atio
n (m
M)
4.50 5.00 5.50 6.00 6.50
75.00
112.50
150.00
120110
100
90
80
70
60
40
30
20 0
80
90
100
10
50
37.50
0.00
0
20
40
60
80
100
120
140
160
0 20 40 60 80 100 120 140 160
Salt concentration (mM)
Conductivity (mS/cm)
10 12 14 1686420
pH 4.5 pH 5.0
100 cm/H300 cm/H700 cm/H
0
1000
2000
3000
4000
5000
6000
mAU
2500
2000
1500
1000
500
30.0 40.0 50.0 60.0 70.0 ml
0
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
500
1000
1500
2000
2500
401
POROS® HS POROS® XS
Volume (mL)
Volume (mL) Volume (mL)Volume (mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Abso
rban
ce a
t 280
nm
(mAU
)
6000
4000
5000
3000
2000
1000
50 10 15 20 25 30 35 40
0
Abso
rban
ce a
t 280
nm
(mAU
)
Capa
city
at 5
% b
reak
thro
ugh
(mg/
mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Salt Concentration (mM)
pH
• Design Points
Salt
Conc
entr
atio
n (m
M)
Figure 4. Resolution as a function of protein loading. Column: 0.46 cmD x 20 cmL; Gradient: 10–100% B, 16 CV; Buffer A: 20 mM MES, 25 mM NaCl pH 6.2; Buffer B: 20 mM Tris, 1.0 M NaCl, pH 6.2Sample: Chymotrypsinogen, Cytochrome C, Lysozyme
Figure 5. IgG binding capacity vs. salt concentration. Column: 0.46 cm x 20 cmL; Buffer: 20 mM MESLoad conditions: 5 mg/mL IgG; Flow rate: 300 cm/h
State-of-the-art manufacturing capability
Life Technologies manufactures POROS® XS resin from approved raw materials to finished product using advanced, proprietary processes that make us unique in this industry. A 30,000 square foot, state-of-the-art chemical facility, in Bedford, Massachusetts, is dedicated to the production of all the POROS® chromatography resins. Quality testing is performed at all stages of the POROS® XS process to ensure excellent lot-to-lot consistency. POROS® XS is backed up with both a drug master file and regulatory support package.
POROS® chromatography resins are manufactured under iSO 9001:2008 and iSO 13485:2003 standards. The facility and quality systems are routinely audited by regulated biopharmaceutical companies.
Conclusion
POROS® XS resin sets the bar in fulfilling the most important demands of today’s high-titer downstream purification processes. This breakthrough cation exchange chromatography resin provides a unique combination of high capacity, high resolution, and high salt tolerance.
The market-leading performance from this dynamic feature set surpasses that of other commercially available cation exchange resins. POROS® XS resin helps enable improved product yield, greater process flexibility, and streamlined downstream processing.
4.50 5.00 5.50 6.00 6.50
75.00
112.50
150.00
120110
100
90
80
70
60
40
30
20 0
80
90
100
10
50
37.50
0.00
0
20
40
60
80
100
120
140
160
0 20 40 60 80 100 120 140 160
Salt concentration (mM)
Conductivity (mS/cm)
10 12 14 1686420
pH 4.5 pH 5.0
100 cm/H300 cm/H700 cm/H
0
1000
2000
3000
4000
5000
6000
mAU
2500
2000
1500
1000
500
30.0 40.0 50.0 60.0 70.0 ml
0
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
500
1000
1500
2000
2500
401
POROS® HS POROS® XS
Volume (mL)
Volume (mL) Volume (mL)Volume (mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Abso
rban
ce a
t 280
nm
(mAU
)
6000
4000
5000
3000
2000
1000
50 10 15 20 25 30 35 40
0
Abso
rban
ce a
t 280
nm
(mAU
)
Capa
city
at 5
% b
reak
thro
ugh
(mg/
mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Salt Concentration (mM)
pH
• Design Points
Salt
Conc
entr
atio
n (m
M)
4.50 5.00 5.50 6.00 6.50
75.00
112.50
150.00
120110
100
90
80
70
60
40
30
20 0
80
90
100
10
50
37.50
0.00
0
20
40
60
80
100
120
140
160
0 20 40 60 80 100 120 140 160
Salt concentration (mM)
Conductivity (mS/cm)
10 12 14 1686420
pH 4.5 pH 5.0
100 cm/H300 cm/H700 cm/H
0
1000
2000
3000
4000
5000
6000
mAU
2500
2000
1500
1000
500
30.0 40.0 50.0 60.0 70.0 ml
0
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
5.2 mg (1.57 mg/mL)
51.8 mg (15.60 mg/mL)
15.5 mg (4.67 mg/mL)
500
1000
1500
2000
2500
401
POROS® HS POROS® XS
Volume (mL)
Volume (mL) Volume (mL)Volume (mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Abso
rban
ce a
t 280
nm
(mAU
)
6000
4000
5000
3000
2000
1000
50 10 15 20 25 30 35 40
0
Abso
rban
ce a
t 280
nm
(mAU
)
Capa
city
at 5
% b
reak
thro
ugh
(mg/
mL)
Abso
rban
ce a
t 280
nm
(mAU
)
Salt Concentration (mM)
pH
• Design Points
Salt
Conc
entr
atio
n (m
M)
HighCapacity
HighResolution
HighSalt Tolerance
XS
Headquarters 850 Lincoln Centre Drive | Foster City, CA 94404 USAPhone 650.638.5800 | Toll Free 800.327.3002 www.appliedbiosystems.com
International Sales For our office locations please call the division headquarters or refer to our website at www.appliedbiosystems.com/about/offices.cfm
For Research Use Only. Not for use in diagnostic procedures.
© 2010 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Printed in the USA. CO21707 1210
Ordering information
Description Pack Size Part Number
POROS® XS Cation Exchange Resin 25 mL 4404339
POROS® XS Cation Exchange Resin 50 mL 4404338
POROS® XS Cation Exchange Resin 250 mL 4404337
POROS® XS Cation Exchange Resin 1,000 mL 4404336
POROS® XS Cation Exchange Resin 5,000 mL 4404335
POROS® XS Cation Exchange Resin 10,000 mL 4404334
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