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Alkali Resistant and Optimized Ligand for High Capacity ... · Semi-rigid Bead for High...
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Alkali Resistant and Optimized Ligand for High Capacity Surface Modified Base Bead for High Purity Semi-rigid Bead for High Productivity
Transcript of Alkali Resistant and Optimized Ligand for High Capacity ... · Semi-rigid Bead for High...
Alkali Resistant and Optimized Ligand for High Capacity
Surface Modified Base Bead for High Purity
Semi-rigid Bead for High Productivity
P-A
P-AP-A
PROTEIN A LIGAND
• High DBC via controlled conformation and orientation
• High alkaline stability from protein engineering
SURFACE MODIFICATION AND BASE BEAD FORMULATION
• Low HCP levels by surface hydrophilization
• High DBC at high flow rate
• Good pressure and flow properties via rigid crosslinking
True polymer innovation that bridges amorphous materials and agarose for advanced protein separation.Protein A chromatography is widely used as the affinity capture step of both mAbs
and Fc-fusion proteins because of its high degree of selectivity. Variations in elution
behavior of the protein A capture step requires more process development work
and could have an impact on the polishing step in the downstream process. Thus
minimizing the variation in elution pH between different molecules for example,
makes it easier to platform the capture process.
PAGE 1
Outstanding Dynamic Binding Capacity• 20% - 50% higher DBC compared to market standard product
Impurity Clearance at Industry Standard• HCP, aggregate, DNA and virus removal are in-line with market standard product
Superior Caustic Stability• More than 100 cycles with 15 mins and 0.1 N NaOH CIP show >90% DBC maintained
Good Pressure Flow Properties• 400 cm/hr at < 3 bar (20 cm BH, 30 cm diameter column)
Attractive Economics• Thanks to a higher productivity and attractive pricing, Protein A media cost can be
reduced by up to 50% compared to market standard product
Amsphere A3
Agarose L
Polymer H
DBC @2 min (g/L)
DBC @4 min (g/L)
DBC @6.6 min (g/L)
HCP (ppm)
BackPressure
(MPa)
AlkalineStability
(%)
40
20
60
55
2000
80
60
40
50
45
35
0
406000
30
0.32
0.16
4000
Summary ofAmsphereTM A3Features
PAGE 2
Dynamic Binding Capacity
0
10
20
30
40
50
mAb1
DB
C (
g/L)
60
mAb2 mAb3 mAb4
mAbs Dynamic Binding Capacities at 5 Minutes
Average Dynamic Binding Capacities of 23 Different mAbs
CONCLUSION:High binding capacities at all flow rates
HPC Clearance
MOLECULE HCCF ELUATE LRV
mAb1
mAb2
mAb3
mAb4
mAb5
350,000
220,000
315,000
1,170,000
560,000
3,100
1,700
3,850
1,650
2,950
2.05
2.1
1.91
2.85
2.27
*Detection by HCP ELISA (Cygnus F550)
10
% B
T D
BC
(m
g/L)
20
40
60
00 2 10 12 16144 6 8
Residence Time (min.)
80
100
120
Average DBC
PAGE 3
10
% B
T D
BC
(g/
L)
10
20
30
40
50
0
Yiel
d (%
)
Run No.
0 10 50 60 90 100807020 30 40
Run No.
60
70
80
90
100
500 40 50 80 90706010 20 30 100
60
70
HC
P L
og R
emov
al V
alue
1
2
3
4
00 10 50 60 90 100807020 30 40
Run No.
DN
A L
og R
emov
al V
alue
1
2
3
4
00 10 50 60 90 100807020 30 40
Run No.
DBC is maintained at initial levels for 100 CIP cycles and Yieldis above 90% and constant for 100 CIP cycles
Dynamic Binding Capacity and mAb Recovery
HCP and DNA Reduction
Protein A Leaching and pH of Eluate
CIP 0.1M NaOH: contact time 30 min0.5M NaOH: contact time 10 min every 10 cycles
Alkaline Stability - CIP CyclingLe
ache
d P
rote
in A
/IgG
[pp
m]
25
30
15
0 10060
Run No.
20
5
10
20 40 80
pH o
f E
luat
e
3
4
5
6
20 10 50 60 90 100807020 30 40
Run No.
5.5
4.5
3.5
2.5
HCP reduction is approximately 2.0 LRV and is constant for 100 CIP cycles;DNA LRV is consistent at approximately 2.9 throughout the 100 CIP cycles
Leached protein A drops to 10-20 ppm after 100 CIPThere are some points under 15ppm; Elution pool pH as a function of run numberis stable at approximately 4.0 - 4.5
10
% B
T D
BC
(g/
L)
10
20
30
40
50
0
Yiel
d (%
)
Run No.
0 10 50 60 90 100807020 30 40
Run No.
60
70
80
90
100
500 40 50 80 90706010 20 30 100
60
70
PAGE 4
0
10
20
30
40
50
mAb1
60
70
mAb2 mAb3 mAb4 mAb5
Leac
hed
PrA
(pp
m/I
gG)
*Detection by Protein A ELISA (Cygnus F740)
Protein A Leaching
Line
ar f
low
vel
ocit
y (c
m/h
r)
100
300
Packed bed height (cm)
10 12 16 18 22
0
200
600
2014
500
800
700
400
24
Packed bed pressure = 3 barPacked bed pressure = 2.5 barPacked bed pressure = 2 bar
CF 1.25
Line
ar f
low
vel
ocit
y (c
m/h
r)
100
300
Packed bed height (cm)
10 12 16 18 22
0
200
600
2014
500
800
700
400
24
Packed bed pressure = 3 barPacked bed pressure = 2.5 barPacked bed pressure = 2 bar
CF 1.30
2 min residence time
3 min residence time
4 min residence time
6 min residence time
2 min residence time
3 min residence time
4 min residence time
6 min residence time
Pressure-Flow Properties
Operating regions(20cm column i.d., CF 1.25)
Operating regions(20cm column i.d., CF 1.30)
Product name AmsphereTM A3
Matrix Methacrylic polymer
Average particle size 50 µm
Ligand Recombinant protein A
Dynamic binding capacity*1 Approximately 54 mg/mL for polyclonal IgG
Maximum operating pressure 0.8 MPa*2
Maximum operating velocity 1200 cm/h (dependent on column size)
Recommended bed height 5 - 25 cm
Working pH range 1 - 13
Cleaning-in-Place stability 0.1 - 0.5 M NaOH
Recommended storage buffer 20 mM sodium phosphate buffer containing 16% Ethanol, pH 7.5
Description
*1 Determined at 10% breakthrough under linear velocity of 300 cm/h in a column with bed height of 20 cm.*2 Do not exceed the column’s pressure resistance.
Technical Properties
Tailored Protein A ELISA kit is available fromCygnus Technologies
Protein A Mix-N-Go ELISA kitfor JSR Life Sciences Ligand (Catalog # F740)
http://www.cygnustechnologies.com/product_detail/protein-a-mix-n-go-elisa-kit-jsr.html
Request yourfree sample by
sending an email to your sales agent.(see contact details
on back page)
Protein A Mix-N-Go ELISA kit
Amsphere™ is a global trademark of JSR Corporation
©2017-2018 JSR Life Sciences – All rights reserved
For more information visit www.jsrlifesciences.com
NORTH AMERICA
JSR Life Sciences JSR Micro, Inc.1280 North Mathilda Avenue Sunnyvale, CA 94089USA +1-408-543-8800 [email protected]
ASIA
JSR Life SciencesJSR Corporation1-9-2 Higashi-Shimbashi,Minato-ku, Tokyo, 105-8640Japan [email protected]
EUROPE
JSR Life Sciences JSR Micro NV Technologielaan 8 B-3001 Leuven Belgium +32-16-668 [email protected]
