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2011 Mitsubishi Chemical Corporation All Rights Reserved.
Chromatographic Media for Antibody / Protein Purification
Separation Materials Department
Mitsubishi Chemical Corporation (MCC)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Technology to control physical properties, such as pore radius, distribution, functionality and surface area
Variety of Installation Records Installed volume >2,200m3 (export 1,400m3, domestic 800m3, in past 5 yrs) Customers: Major pharmaceutical companies (in JPN, US, India, UK, Germany, FR, Switzerland, Ireland, etc)
Major applications:
Human Insulin Lisinopril Cephalosporin C Vancomycin Daptomycin Cefotaxime Imipenem
New: Antibody/ Protein
P.2
-
100,000
200,000
300,000
400,000
500,000
2007 2008 2009 2010 2011 2012
Export Domestic Total
Vo
lum
e (L)
year
Background: MCC Bioseparation Media
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.3
Peptides, Proteins MW 1000-100,000 Macrolides
MW 50- 1500
10 100 1000Pore size ()
SP825L HP21
SP850
SP70 SP700
HP20
SP207
-Lactums MW ~600
HP2MGL
600-800
1,000-2,000
4,000-5,000
(Reference) Ability to control media properties
Example) Synthetic Adsorbent SEPABEADSTM
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.4
Protein A affinity chromatography
MabSpeedTM rP series
Ion-exchange chromatography
ChromSpeedTM series
type S strong cation exchanger
type Q strong anion exchanger
type CM weak cation exchanger
type DA weak anion exchanger
--- New Media for Biopharmaceutical industry --- Highly productive & efficient purification via high throughput!
NEW ADDITION to the family!!!
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.5
--- New Production Facility for Bio-separation Media ---
Built in February 2013
Prepared in clean dedicated facility
Hygiene management by ISO9001
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.6
--- Product Lineup for MCC Bio-separation Media---
Affinity Ion Exchange
Product MabSpeedTM ChromSpeedTM
rP S Q CM DA
Matrix Crosslinked Polymethacrylate
Ligand / Functionality
rProtein-A -SO3- -N+(CH3)3 -COO
- -N+H(CH3)2
Particle Size (m)
35, 45 30, 60
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.7
Spherical and mono-disperse particles
Easy and reproducible packing
Low pressure drop
Non-compressive bed
Rigid and durable matrix
No volume change
Longer life
Wide pH & solvent compatibility
MCCs New Bio-separation Media---
High performance for achieve high throughput purification
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Production/Sales
Mitsubishi Chemical Co. (Japan)
Tai Young Chemical Co., Ltd.
Resindion (Italy)
Sales Mitsubishi Chemical China Commerce Ltd. Mitsubishi Chemical India Pvt. Ltd. ITOCHU Chemicals America Inc.
Resindion
TYC
MCC ICAI MCN
MCI
Sales/Production network
SYFT
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Affinity chromatography MabSpeedTM
With wild type ligand: MabSpeedTM P102 (45 m) & rP111 (35 m) With engineered type ligand: MabSpeedTM P202 (45 m)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.10
--- MCC Protein-A affinity media line-up--
Mode Protein-A Affinity
Product MabSpeedTM
rP102 rP111 rP202*
Matrix Crosslinked Polymethacrylate
Ligand / Functionality
rProtein-A
(wild type)
rProtein-A
(engineered type)
Particle Size (m) 45 35 45
DBC (g/L-resin)
10% breakthrough,
-globulin, r.t. 3min)
24 33 >50
*Matrix of MabSpeedTM rP202 has been modified to have even higher DBC with the original ligand. *MabSpeedTM rP202 media/columns are currently avilable in Asia and will be available in other regions in the end of 2016. For details, please inquire us.
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.11
:*Pressure drop data. Data was taken with a condition of a column 20 ID x 200 mm with liquid water at temperature 25 C.
--- Hydraulic data represents low pressure drop & easy packing ---
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.12
MabSpeedTM 0 G MabSpeedTM 5,000 G
Polysaccharide 0 G Polysaccharide 5,000 G
reversible
irreversible
Pressure durability
2014 Mitsubishi Chemical Corporation All Rights Reserved.
600
GE MabSelect SuRe
200
400
200 400
200
MabSpeedTM rP102
MabSpeedTM rP111
DBC (mg-IgG/mL-resin)
A2
80
(C/C
0, %
) Breakthrough profiles at various flow rate
(cm/h) 200 400 600 MabSelect SuRe
42.3 32.7 -
MabSpeedTM
rP111 35.6 32.5 30.4
MabSpeedTM
rP102 26.1 24.0 16.2
Dynamic binding capacity
Column 5mm ID x 200 mm Sample: 1.0 mg/mL human -globulin Buffer: PBS pH 7.4, Temp: 20 deg C The DBC was determined at 10% breakthrough. Flow: 200, 400, 600 cm/h
(10% BTC)
Fast kinetics, both fast adsorption and desorption (later discussed) leads to shorter operation time; high IgG production!
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Wild type ligand: MabSpeedTM P102 (45 m)
& MabSpeedTM rP111 (35 m)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
mAb purification from cell culture
Conditions: Column: MabSpeed rP111 5mm I.D. x 20cm. (BV:4.0mL ) Binding buffer PBS Wash buffer PBS Elution buffer 0.1M Citrate pH3.0 Flow rate 400cm/h System AKTA avant Sample CHO cell culture containing 0.1mg/mL mAb, 100mL
mL
A2
80
>HCP contamination Start material: 116,160 ppm(ng-HCP/mg-IgG) (HCP: 11,616 ng/mL IgG: 0.1mg/mL) IgG fraction - MabSpeedTM rP111: 9.2 ppm(ng-HCP/mg-IgG) - MabSelect SuRe 63.7 ppm >PrA ligand leakage - MabSpeedTM rP111: 3.4 ppm (ng-PrA/mg-IgG) >Recovery - >98% (UV280nm)
1 Molecular weight marker 2 start material 3 path through 4 mAb fraction
1 2 3 4
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.16
CIP durability
DB
C(%
) P
rA leak (p
pm
)
5.62
0.25
cycles Column: 5mm ID x 5cm Binding: Phsphate bufferd saline(pH7.4) Elution: 0.1M Sodium Citrate pH3.0 CIP: 0.1M NaOH Contact: 15min
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.17
@ pH 3.0
@ pH3.5
MabSpeedTM
(rProtein A) Other polymer media
rP102 data
Spin column 0.5mL resin single cycle (1-6)
1. equilibration buffer A, 6 BV
2. sample load 1mg/mL g-globulin, 2 BV
3. washout buffer A, 6 BV
4. elution buffer B, 4 BV
5. regeneration buffer C, 4 BV
Each Step 500 g x 1 min
buffer A: PBS pH7.4
buffer B: 0.1M sodium citrate pH3.0, 3.5
buffer C: 1M Tris-HCl pH9.0
Agarose (alkali resistant)
Effect of elution pH
- Sharp elution for MabSpeed at pH 3.5 - Elution conditions for wild type of rProtein-A ligand can be applied for MabSpeed.
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.18
Faster equilibrium of MabSpeed contributes buffer and time savings about 30%
Buffer exchanging profiles
MabSpeedTM rP111 MabSelect SuRe
200 400
100 200 400
100 Column : 10 ID x 200 mm Base : 0.8M NaCl Load : 0.05M NaCl, Temp : 20 deg-C Flow : 100, 200, 400 cm/h
(cm/h) 100 200 400
MabSpeedTM rP111 10.7 5.4 2.8
MabSelect SuRe 14.0 7.2 3.8
Time to equilibration (min, ~6.0>mS/cm)
Flow time (min)
Co
nd
uct
ivit
y(m
S/cm
)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
MabSelect SuRe
400cm/h
min/cycle
DBC (1% BTC,g/L-resin)
cycle/day
(adsorption) (misc)
IgG Production (g/day)
62 60
11.8
20.6
382
Colume volume (L) 1.57
flow rate (cm/h)
Column height (cm)
Productivity Simulation
MabSpeed rP111
400cm/h 600cm/h
89 48
10.5
29.8
56 37
15.4
28.2
489 681
1.57 1.57
200cm/h
200 105
4.7
33.3
247
1.57
200cm/h
206 81
5.0
34.4
270
1.57
MabSpeedTM rP111 results in higher productivity than MabSelect SuRe
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.20
Mab purification Chromatography with MCC resins
Protein A affinity : MabSpeedTM rP111
Cation exchange : ChromSpeedTM S101
Anion exchange : ChromSpeedTM Q101
Column MabSpeedTM rP111 10x150mmH(BV: 12mL) Sample Clarified CHO cell culture, 0.5mg/mL mab (20.8mg/mL-resin) Equiliburation buffer 20mM Sodium Phosphate, 150mM NaCl, pH7.4 Elution buffer 20mM Sodium Citrate, pH3.4 Flow 300 cm/h, R.T. 3min Temp. 20 deg-C
Column ChromSpeedTM S101 5x200mmH(BV: 4mL) Sample mab (51.3 mg/mL-resin) after purification on MabSpeedTM rP111 Start buffer 20mM Sodium acetate, pH5.5 Elution buffer 20mM Sodium acetate, 1M NaCl, pH5.5 Flow 300 cm/h, R.T. 4min Gradient 5% (10CV), 15% (20CV), 100% (10CV) Temp. 20 deg-C
Column ChromSpeedTM Q101 5x50mmH(BV: 1mL) Sample mab (192 mg/mL-resin), eluate after ChromSpeedTM S101, pH adjusted to 6.0 Start buffer 20mM Sodium acetate, pH6.0 Flow 300 cm/h, R.T. 1min Temp. 20 deg-C
capture, removal of aggregate, HCP,
removal of agg, HCP, PrA
removal of DNA, endotoxin,
Cell culture
Three step processAccumulated
yield (%)
Dimers and
aggregate (%)
Protein A
(ppm)
HCP
(ppm)
Start material 100 - - 35717
MabSpeedTM rP111 98
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Engineered type ligand: MabSpeedTM P202 (45 m)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
High Dynamic Binding Capacity (DBC):
P.22
DBC of MabSpeed rP202, well exceeding those agarose and/ or polymeric media in standard flow rate in 100 cm/hr (r.t of
12 min) to 400 cm/hr.
DBC of MabSpeed rP202: keeps the high DBC even at high folwrate in the range of 600 cm/hr (r.t. of 2 min) to 1,000 cm/hr (r.t. of 1.2
min)
Drawback of agarose media: high pressure drop
(or back pressure) prevents to utilize high flow rates, sometimes resulting very low DBC.
R.T.(min) 1.5 2.0 3.0 6.0
1 g/L IgG 42 44 51 62
5 g/L IgG 38 44 50 59
MabSpeed rP202 DBC
*Data was taken with a column 5 x 200mmH with a sample of 1 mg/mL human IgG. The straight red arrow shows high DBC at high flow rate while the blank green arrow of agarose media shows significant decrease of DBC.
2014 Mitsubishi Chemical Corporation All Rights Reserved.
CIP Durability
P.23
Cycles
DB
C (
%)
Lig
an
d le
ak
(pp
m)
*Experiments were performed with a column of 5mm ID x 5cm, by binding with phosphate buffered saline(pH7.4) and eluting with 0.1M sodium citrate at pH3.0. The contact time was 15min.
High DBC and Protein-A ligand leakage (
2014 Mitsubishi Chemical Corporation All Rights Reserved.
MabSpeed rP202 new engineered ligand enables
elution with weakly acidic condition of pH 4.0:
P.24
3.9 3.4 3.9
3.9
Numbers in the profiles are pH of eluate pool. Data are taken with a condition of a column 5 x 50 mmH (1mL), flow rate 1.0 mL/min, eluent A 0.1 M, citrate pH6.5, eluent B 0.1 M citrate pH2.5, pH gradient 0->100 %B over 20 CV (20 mL), and with a sample of 20 g/mL mab in TST, 10 mL (0.2 mg):
MabSpeedTM rP111 (wild type ligand)
MabSpeedTM rP202 (engineered type ligand)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
CHO cell culture purification
P.25
Low HCP contamination and Protein-A ligand leakage with high recovery
A2
80
mL
>HCP contamination Start material: 116,000 ppm(ng-HCP/mg-IgG)
IgG fraction : 19 ppm(ng-HCP/mg-IgG) (3.8 log down) >Protein A ligand leakage IgG fraction : 6.3 ppm (ng/mg-IgG)
Conditions: Column MabSpeed rP202 26 x 7mm I.D. (BV:1.0mL ) Binding buffer PBS Wash buffer PBS Elution buffer 0.1M Citrate pH3.0 Flow rate 1.0 mL/min System AKTA avant Sample CHO cell culture containing 0.99 mg/mL mAb, 30mL
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Buffer Exchange
P.26
Flow time (min)
Co
nd
uct
ivit
y(m
S/cm
)
200 cm/hr
400 cm/hr
100 cm/hr
(cm/h) 100 200 400 600
MabSpeedTM rP202
17.1 8.6 4.4 3.0
MabSelect SuRe
18.8 9.8 5.0 X
Time to equilibration (min, ~6.0>mS/cm)
Column : 10 ID x 200 mm Base : 0.8M NaCl Load : 0.05M NaCl, Temp : 20 deg-C Flow : 100, 200, 400 600cm/h
~ 10% saving of buffers due to fast desorption with MabSpeedTM rP202!
2014 Mitsubishi Chemical Corporation All Rights Reserved.
MabSelect SuRe
400cm/h
min/cycle
DBC (1% BTC,g/L-resin)
cycle/day
(adsorption) (misc.)
IgG production (g/day)
98 60
9.1
33
468
Column volume (L) 1.57
flow rate (cm/h)
Column height 20cm
Productivity simulation 1
MabSpeed rP202
400cm/h 600cm/h
153 54
6.9
51
88 42
11.1
44
555 769
1.57 1.57
200cm/h
254 105
4.0
42
267
1.57
200cm/h
372 93
3.1
62
301
1.57
IgG production: MabSpeedTM rP202 well exceeds MabSelect SuRe
2014 Mitsubishi Chemical Corporation All Rights Reserved.
MabSelect SuRe LX
400cm/h
121 60
7.9
40
504
1.57
MabSpeed rP202
400cm/h 600cm/h
153 54
6.9
51
88 42
11.1
44
555 769
1.57 1.57
200cm/h
326 105
3.3
54
285
1.57
200cm/h
372 93
3.1
62
301
1.57
Productivity simulation 2
min/cycle
DBC (1% BTC,g/L-resin)
cycle/day
IgG production (g/day)
Column volume (L)
flow rate (cm/h)
Column height 20cm
IgG production: MabSpeedTM rP202 well exceeds MabSelect SuRe LX
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Ion exchange chromatography ChromSpeedTM
ChromSpeedTM S103, Q103, CM103, and DA103 (60 m) ChromSpeedTM S101, Q101, CM101, and DA101 (30 m)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.30
Batch adsorption capacity
Ion exchange capacity
(eq/L-resin)
Batch adsorption capacity
(g/L-resin)
Human -globulin
0.09 125
Functional group
60
Particle diameter
(m)
124 0.08 60
SBC condition: S- and CM- type: Human -globulin 2.5 g/L, 20 mM citrate (pH 5.2), 25 Q- and DA- type Human -globulin 2.5 g/L, 20 mM Tris-HCl (pH 9.0), 25
-N+H(CH3)2
-COO 114 0.11 60
81 0.13 60
0.08 140 30
130 0.08
-N (CH3)3
30
98 0.11 30
120 0.10 30
-SO3
ChromSpeed
S103
S101
Q103
Q101
CM103
CM101
DA103
DA101
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.31
Pressure drop comparison (Water, 25C)
0.00
0.40
0.80
1.20
1.60
2.00
0 500 1000 1500 2000
Linear velocity (cm/h)
Pre
ssu
re d
rop
(M
Pa
/m)
ChromSpeed Q103, 60m
Agarose-based resin 1, 90m
Agarose-based resin 2, 90m
Excellent Hydraulic Property --- No bed compression at 1,800cm/hr linear velocity ---
Hydraulic properties of ChromSpeedTM
2014 Mitsubishi Chemical Corporation All Rights Reserved.
1-1) S103 (60m) vs competitors
ChromSpeedTM S103 (Mitsubishi)
UV
28
0n
m
Capto S (GE)
(a) (b) (a) (b)
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
Gigacap S 650M (TOSOH)
(a) (b)
60m 90m 50-100m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Effect of IgG Concentration
P.33
Column 5 x 100 mmH (BV:2.0mL) Buffer 20mM Sodium Acetate (pH5.5) Flow rate 200, 400, 600, 800 cm/h (R.T. 3.0, 1.5, 1.0, 0.75 min) Sample a) 1.0, b) 2.5, c) 10 mg/mL gamma-globulin (IgG)
a) 1.0 mg/mL b) 2.5 mg/mL
10
% D
BC
(m
g-Ig
G/m
L-re
sin
)
c) 10 mg/mL
The highest dynamic binding capacity was observed for ChromSpeedTM S103. Moreover, DBC showed less dependence on the IgG concentration for ChromSpeedTM S103.
2015 Mitsubishi Chemical Corporation All Rights Reserved.
Durability of ChromSpeedTM S103
SBC *1 >120 mg-IgG/mL-resin Recovery*2 >99% (IgG) Pressure Drop*3 < 1.5MPa/m at 1,000 cm/h Alkaline tolerance*4 100cycles of 0.5M NaOH exposure O.K.
*1 2.5 g/L, 20 mM Sodium acetate (pH 5.5), 20 deg-C *2 1.0 g/L, 20 mM Sodium acetate (pH 5.5), 20 deg-C *3 10x200mmH, 150mM NaCl, room temperature *4 0.5M NaOH, 15min/cycle, 20deg-C, DBC: IgG at 10 % breakthrough
cycles
DB
C(%
)
2014 Mitsubishi Chemical Corporation All Rights Reserved.
ChromSpeedTM S103 (Mitsubishi)
UV
28
0n
m
(a) (b)
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
ChromSpeedTM S101 (Mitsubishi)
(a) (b)
1-2) S103 (60m) vs S101 (30m)
60m 30m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
ChromSpeedTM S101 (Mitsubishi)
Capto SP ImpRes (GE)
UV
28
0n
m
(a) (b)
(a) (b)
1-3) S101 (30m) vs competitors
36-44m 30m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
ChromSpeedTM Q103 (Mitsubishi)
UV
28
0n
m
Gigacap Q (TOSOH)
(a)
(b)
(min) (min)
(a)
(b)
Conditions: Column 100 x 5mm I.D. (BV 2mL) Eluent A 50mM Tris-HCl (pH8.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Myoglobin + (b) Trypsin inhibitor = 250/50ug / 25uL (pI) (7.5) (4.5)
2-1) Q103 (60m) vs competitors
60m 50-100m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
ChromSpeedTM Q103 (Mitsubishi)
UV
28
0n
m
(a)
(b)
Conditions: Column 100 x 5mm I.D. (BV 2mL) Eluent A 50mM Tris-HCl (pH8.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Myoglobin + (b) Trypsin inhibitor = 250/50ug / 25uL (pI) (7.5) (4.5)
(min) (min)
(a)
(b)
ChromSpeedTM Q101 (Mitsubishi)
2-2) Q103 (60m) vs Q101 (30m)
60m 30m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Capto Q ImpRes (GE)
UV
28
0n
m
Conditions: Column 100 x 5mm I.D. (BV 2mL) Eluent A 50mM Tris-HCl (pH8.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Myoglobin + (b) Trypsin inhibitor = 250/50ug / 25uL (pI) (7.5) (4.5)
(min) (min)
(a)
(b)
2-3) Q101 (30m) vs competitors
(min)
(a)
(b)
ChromSpeedTM Q101 (Mitsubishi)
30m 36-44m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
UV
28
0n
m
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
GigaCap CM 650M (TOSOH)
(a) (b)
ChromSpeedTM CM103 (Mitsubishi)
(a) (b)
3-1) CM103 (60m) vs competitors
Comparable product unavailable
Capto CM (GE)
UV
28
0n
m
60m 60m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
UV
28
0n
m
(a) (b)
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
ChromSpeedTM CM101 (Mitsubishi)
ChromSpeedTM CM103 (Mitsubishi)
(a) (b)
3-2) CM103 (60m) vs CM101 (30m)
UV
28
0n
m
60m 30m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
P.42
4-1) DA103 (60m) vs competitors
ChromSpeed-DA103 Agarose-based DEAE medium
Conditions: Column, 100 x 9mmI.D. (6.4ml); Conditions: Column, 5ml;
Black Eluent A, 20mM sodium phosphate (pH7.0); Black Eluent A, 20mM sodium phosphate (pH7.0);
Eluent B, A + 1.0M NaCl; Eluent B, A + 1.0M NaCl;
Blue Eluent A, 20mM Glycine-NaOH (pH10.0); Blue Eluent A, 20mM Glycine-NaOH (pH10.0);
Eluent B, A + 1.0M NaCl; Eluent B, A + 1.0M NaCl;
Flow rate, 1.0ml/min (94cm/h); Gradient, 0-50% B over 30min. Flow rate, 0.786ml/min; Gradient, 0-50% B over 30min.
Samples: a-Lactalbumin, 160mg / 160ml Samples: a-Lactalbumin, 125mg / 125ml
-9.00E+03
0.00E+00
9.00E+03
1.80E+04
2.70E+04
3.60E+04
4.50E+04
5.40E+04
6.30E+04
7.20E+04
8.10E+04
0 5 10 15 20 25 30 35 40
time (min)
UV
28
0n
m (m
V)
-9.00E+03
0.00E+00
9.00E+03
1.80E+04
2.70E+04
3.60E+04
4.50E+04
5.40E+04
6.30E+04
7.20E+04
8.10E+04
0 5 10 15 20 25 30 35 40
time (min)
UV
28
0n
m (m
V)
pH7pH10
pH10pH7
Capto DEAE (GE)
ChromSpeed DA103 (Mitsubishi)
60m 90m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
UV
28
0n
m
ChromSpeedTM DA101 (Mitsubishi)
ChromSpeedTM DA103 (Mitsubishi)
4-2) DA103 (60m) vs DA101 (30m)
Conditions: Column, 50 x 5mmI.D. (60mm); Eluent A, 20mM sodium phosphate (pH7.0); Eluent B, A + 1.0M NaCl; Flow rate, 0.5ml/min (150cm/h); Gradient, 0-50% B over 30min. Samples: a-Lactalbumin / Trypsin inhibitor = 25mg / 50mg / 25ml
60m 30m
2014 Mitsubishi Chemical Corporation All Rights Reserved.
Product Details
Standard package size: 25 / 100 / 1000 mL Slurry in 20%-Ethanol
Documentations
COA MSDS RSF (NDA required)
Screening columns available for ChromSpeedTM 103 Series 1 mL columns 5 mL columns
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Thank you
2011 Mitsubishi Chemical Corporation All Rights Reserved.
Reference
2015 Mitsubishi Chemical Corporation All Rights Reserved.
ChromSpeedTM S103 (60um, Mitsubishi)
ChromSpeedTM S101 (30um, Mitsubishi)
Capto S (90um, GE healthcare)
Capto SP ImpRes (40um, GE healthcare)
GigaCap S650M (50-100um, TOSOH)
CellufineMax S (40-130 um, JNC)
Nuvia S (85um, Bio-Rad)
UV
28
0
time(min)
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 20mM Sodium Phosphate (pH6.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample (a) Cytochrome C + (b) lysozyme = 125 / 125ug / 50uL (pI) (9.3) (11.0)
GigaCap S650S (20-50um, TOSOH)
SP Sepharose FF (90um, GE healthcare)
BioPro S75 (75um, YMC)
Eshmuno S (75-95um, Merck Millipore)
-50000
0
50000
100000
150000
200000
250000
300000
350000
400000
450000
500000
0 10 20 30 40
Selectivity of Strong Cation Exchangers
2015 Mitsubishi Chemical Corporation All Rights Reserved.
-10000
0
10000
20000
30000
40000
50000
60000
70000
0 5 10 15 20 25 30 35 40ChromSpeed Q103 (60um, Mitsubishi)
ChromSpeed Q101 (30um, Mitsubishi)
Gigacap Q650M (50-100um, TOSOH)
Gigacap Q650S (20-50um, TOSOH)
Q Sepharose FF (90um, GE healthcare)
Capto Q (90um, GE healthcare)
CaptoQ ImpRes (40um, GE healthcare)
time(min)
Conditions: Column 100 x 5mm I.D. (BV:2.0mL ) Eluent A 50mM Tris-HCl (pH8.5) Eluent B A + 1.0M NaCl Flow rate 1.0mL/min (R.T. 2min) Gradient 0-100% B over 60min Sample alpha lactalbumin + Trypsin inhibitor = 25 / 125ug / 25uL (pI) (4.4) (4.5)
UV
28
0
Selectivity of Strong Anion Exchangers