Download Analysis of Monoclonal Antibody and Related Substances using a

Analysis of Monoclonal Antibody and Related
Substances using a New Hydrophobic Interaction
Chromatography (HIC) Column
Julia Baek,1 Robert van Ling, 2 Xiaodong Liu1
1
Thermo Fisher Scientific, Sunnyvale, CA, USA; 2Thermo Fisher Scientific, Breda, The Netherlands
Overview
FIGURE 1. Fast se
160
Purpose: Demonstrate high resolution separation of mAbs
and mAb related substances using Thermo ScientificTM
MAbPacTM HIC-10 column.
Introduction
Monoclonal antibodies (MAbs) are the most prominent class
ofprotein therapeutics because of their high specificity,
excellent biocompatibility and effectiveness against
autoimmune disorders, cardiovascular diseases, infectious
diseases, and cancer. The proliferation of monoclonal
antibody therapeutics and their susceptibility to various
biochemical modifications has highlighted the importance of
characterizing these highly heterogeneous products for their
safety and efficacy.
Hydrophobic interaction chromatography (HIC) is a
technique for separation of proteins including monoclonal
antibodies. The HIC mobile phase usually consists of a
salting-out agent, which at high salt concentration, retains the
protein by increasing hydrophobic interaction between the
protein and the stationary phase. HIC has been widely used
as an orthogonal method to size exclusion chromatography
and ion exchange chromatography for analysis of mAb and
related substances, such as succinimides, fragments,
oxidants, C-terminal lysine modification, and drug
conjugates, to monitor the stability and potency of the drug.
Here we introduce a new HIC column designed for mAb
analysis. It is based on high-purity, spherical, wide-pore
(1,000 Å), 5 µm silica particles functionalized with proprietary
alkyl amide groups. The advanced surface bonding
technology provides desired selectivity, excellent recovery,
high efficiency, and chemical stability. Its effectiveness for
mAb separation has been demonstrated by examples
including mAb fragments, oxidized mAb, PEGylated mAb,
mAb aggregates and antibody-drug conjugates (ADCs).
2 Analysis of Monoclonal Antibody and Related Substances using a New Hydrophobic Interaction Chromatography (HIC) Column
Absorbance (mAU)
80
40
0
0
2
4
Analysis of mA
Analysis of antibod
characterization of
localization of the s
antibody molecule.
the separation of Fa
hydrophobic varian
mAb and its papain
HIC-10 column effic
further separates va
peaks imply oxidati
FIGURE 2. Separa
a
30
Absorbance (mAU)
Results: The new MAbPac HIC-10 column was developed
using advanced surface bonding technology to achieve
unique selectivity, high recovery and high efficiency. High
resolution separation of proteins and various mAb samples
were successfully carried out using a MAbPac HIC-10
column.
20
10
0
20
Absorbance (mAU)
Methods: Ammonium sulfate and sodium phosphate mobile
phase were used. In some cases ,addition of isopropanol to
both mobile phases increased the resolution of the
chromatogram.
120
b
15
10
5
0
0
2
4
Analysis of Oxi
Monoclonal antibod
storage and deliver
residues have been
mAb therapeutics.2
Here we introduce a new HIC column designed for mAb
analysis. It is based on high-purity, spherical, wide-pore
(1,000 Å), 5 µm silica particles functionalized with proprietary
alkyl amide groups. The advanced surface bonding
technology provides desired selectivity, excellent recovery,
high efficiency, and chemical stability. Its effectiveness for
mAb separation has been demonstrated by examples
including mAb fragments, oxidized mAb, PEGylated mAb,
mAb aggregates and antibody-drug conjugates (ADCs).
10
5
0
0
2
4
Analysis of Oxi
Monoclonal antibod
storage and deliver
residues have been
mAb therapeutics.2
confirm the stability
MAbPac HIC-10 co
from the native mA
Methods
Samples
Monoclonal antibody samples and ADC sample were
donated by biotech companies. Proteins and other chemicals
were from Sigma-Aldrich®.
FIGURE 3. Separa
14
Column
MAbPac HIC-10, 5 µm, 4.6 × 100 mm (P/N 088480)
Absorbance (mAU)
12
Liquid Chromatography
HPLC experiments were carried out using Thermo
Scientific™ Dionex™ UltiMate™ 3000 BioRS system
equipped with:
10
8
6
4
2
 SR-3000 Solvent Rack (PN 5035.9200)
 LPG-3400RS Biocompatible Quaternary Rapid Separation
Pump (PN 5040.0036)
 WPS-3000TBRS Biocompatible Rapid Separation
Thermostatted Autosampler (PN 5841.0020)
 TCC-3000RS Rapid Separation Thermostatted Column
Compartment (PN 5730.0000)
 VWD-3400RS Rapid Separation Variable Wavelength
Detector (VWD) equipped with micro flow cell (PN
5074.0010)
 Chromatography was controlled by Thermo Scientific™
Dionex™ Chromeleon Chromatography Data System.
0
0
5
Re
Analysis PEGyl
PEGylated proteins
biotherapeutics. In
immunogenicity, inc
proteolysis, resultin
desirable to attach
reaction is not alwa
be attached it is imp
analyze the purity a
mAbs were succes
column in 20 minut
ammonium sulfate
the gradient resulte
Method Development
Ammonium sulfate and sodium phosphate based mobile
phases were used. For the applications with mAb and mAb
related substances, mobile phases were optimized by either
lowering the starting salt concentration or adding isopropanol
into both mobile phase A and mobile phase B.
FIGURE 4. Separa
180
Results
150
High Resolution and Fast Separation of Proteins
125
•
bance (mAU)
The MAbPac HIC-10 column is based on high-purity,
Thermo Scientific Poster Note
spherical, wide-pore (1,000 Å), 5 µm silica particles
15
Absorbance (mA
oxidants, C-terminal lysine modification, and drug
conjugates, to monitor the stability and potency of the drug.
100 ISC 0814S 3
PN21024
FIGURE 1. Fast separation of standard protein mixture
Analysis of
160
separation of mAbs
mo ScientificTM
120
Absorbance (mAU)
m phosphate mobile
on of isopropanol to
ution of the
Column:
Format:
Mobile phase A:
4
Mobile phase B:
3
1
80
Gradient:
2
Temperature:
Flow rate:
Inj. volume:
Detection:
Sample:
Peaks:
40
umn was developed
ology to achieve
h efficiency. High
rious mAb samples
AbPac HIC-10
0
0
2
4
6
Retention Time (min)
8
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
2 M ammonium sulfate, 100 mM
sodium phosphate, pH 7.0
100 mM sodium phosphate, pH
7.0
Time (min)
-5.0
0.0
1.0
7.0
10.0
%A
100
100
100
0
0
%B
0
0
0
100
100
30 ºC
1.0 mL/min
10 µL
UV (280 nm)
Protein mixture
1) Myoglobin
2) Ribonuclease A
3) Lysozyme
4) α-Chymotrypsinogen A
Protein and an
product expres
storage. These
reactions whic
widely used te
protein aggreg
researchers ha
protein aggreg
monoclonal an
MAbPac HIC-1
than the main
FIGURE 5. Se
10
300
Analysis of mAb Fragments
esigned for mAb
erical, wide-pore
alized with proprietary
ace bonding
excellent recovery,
s effectiveness for
d by examples
, PEGylated mAb,
jugates (ADCs).
4
FIGURE 2. Separation of papain digested mAb
a
30
Absorbance (mAU)
y (HIC) is a
uding monoclonal
lly consists of a
centration, retains the
action between the
as been widely used
on chromatography
analysis of mAb and
es, fragments,
and drug
potency of the drug.
Column:
Format:
Mobile phase A:
Intact mAb
20
Mobile phase B:
10
Gradient:
0
20
b
Fab
Temperature:
Flow rate:
Inj. volume:
15
10
Detection:
Sample:
Fc
5
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0 /
isopropanol (95:5 v/v)
50 mM sodium phosphate, pH
7.0 / isopropanol (80:20 v/v)
Time (min)
-5.0
0.0
1.0
15.0
20.0
%A
100
100
100
0
0
%B
0
0
0
100
100
25 ºC
1.0 mL/min
Papain digest:10 µL (1 mg/mL)
Intact mAb:
2 µL (5 mg/mL)
UV (280 nm)
a) Intact mAb
b) Papain digest
0
0
2
4
6
Retention Time (min)
8
10
Analysis of Oxidized mAb
Monoclonal antibodies are susceptible to oxidation during
storage and delivery. Oxidation of methionine or tryptophan
residues have been linked to decreased or loss of bioactivity of
Analysis of Monoclonal Antibody and Related Substances using a New Hydrophobic Interaction Chromatography (HIC) Column
mAb therapeutics.2 Therefore it is critical to monitor oxidation to
Absorbance (mAU)
Analysis of antibody fragments is important for both
characterization of Fab or Fc based biotherapeutics and
localization of the sources of heterogeneities on a monoclonal
antibody molecule. HIC can provide the resolution required for
the separation of Fab and Fc fragments and their hydrophilic or
hydrophobic variants.1 Figure 2 shows a comparison of an intact
mAb and its papain digest on MAbPac HIC-10. The MAbPac
HIC-10 column efficiently separates Fab and Fc fragments and
further separates variants of these fragments. These variant
peaks imply oxidation or other modifications in these fragments.
Absorbance (mAU)
most prominent class
high specificity,
ess against
diseases, infectious
of monoclonal
bility to various
ed the importance of
ous products for their
250
200
150
100
50
0
0
2
4
Analysis of
Antibody drug
protein therape
interaction chro
ADCs since att
the antibody. T
elutes first and
elution time of
often used to c
different drug-t
separation of a
MAbPac HIC-1
between a drug
interchain cyst
loaded antibod
unmodified mA
are well resolv
shape and sep
phase A and 20
P/N 088480)
ng Thermo
BioRS system
hermo Scientific™
y Data System.
ate based mobile
with mAb and mAb
e optimized by either
or adding isopropanol
ase B.
ation of Proteins
n high-purity,
ca particles
de groups. First, a
Myoglobin,
otrypsinogen was
hase. A fast 6-minute
6
Retention Time (min)
8
10
Monoclonal antibodies are susceptible to oxidation during
storage and delivery. Oxidation of methionine or tryptophan
residues have been linked to decreased or loss of bioactivity of
mAb therapeutics.2 Therefore it is critical to monitor oxidation to
confirm the stability and clinical efficacy of mAb products.
MAbPac HIC-10 column was able to separate oxidized mAb
from the native mAb as shown in Figure 3.
MAbPac HIC-1
between a drug
interchain cyste
loaded antibod
unmodified mA
are well resolve
shape and sep
phase A and 20
FIGURE 6. Sep
40
FIGURE 3. Separation of oxidized mAb
30
14
Column:
Format:
Mobile phase A:
12
Mobile phase B:
Gradient:
10
8
6
4
Temperature:
Flow rate:
Inj. volume:
2
Detection:
Sample:
0
0
5
10
Retention Time (min)
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
2 M ammonium sulfate, 100 mM
sodium phosphate, pH 7.0
100 mM sodium phosphate, pH 7.0
Time (min)
-5.0
0.0
1.0
15.0
20.0
%A
60
60
60
0
0
%B
40
40
40
100
100
FIGURE 4. Separation of PEGylated mAbs
180
Column:
Format:
Mobile phase A:
150
Mobile phase B:
125
Gradient:
100
75
50
25
2
10
0
0
5
1
Conclusio
15
PEGylated proteins are an important class of modern
biotherapeutics. In general, PEGylation decreases
immunogenicity, increases circulatory time and reduces
proteolysis, resulting in enhanced medical efficacy.3 Typically it is
desirable to attach one PEG at a specific site. However since the
reaction is not always site specific and more than one PEG may
be attached it is important to purify the PEGylated protein and
analyze the purity afterwards. Two derivatives of PEGylated
mAbs were successfully separated using MAbPac HIC-10
column in 20 minutes (Figure 4). Lowering the starting
ammonium sulfate concentration to 0.6 M by simply changing
the gradient resulted in a higher resolution separation.
1
20
30 ºC
1.0 mL/min
mAb:
2 µL (2.5 mg/mL)
Oxidized mAb: 4 µL (1.3 mg/mL)
UV (280 nm)
mAb
Oxidized mAb
Analysis PEGylated mAb
Absorbance (mAU)
ble Wavelength
flow cell (PN
4
Analysis of Oxidized mAb
0)
ry Rapid Separation
Separation
.0020)
mostatted Column
2
Absorbance (mAU)
sample were
s and other chemicals
0
Absorbance (mAU)
lized with proprietary
ce bonding
excellent recovery,
s effectiveness for
d by examples
, PEGylated mAb,
jugates (ADCs).
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0
50 mM sodium phosphate, pH
7.0
Time (min)
-5.0
0.0
1.0
15.0
17.0
17.1
23.0
%A
40
40
40
0
0
40
40
Temperature:
30 ºC
Flow rate:
0.5 mL/min
Inj. volume:
10 µL (1 mg/mL)
Detection:
UV (280 nm)
ThermoPEGylated
Scientific
Poster
Sample:
mAb
Peaks:
1) Derivative 1
2) Derivative 2
%B
60
60
60
100
100
60
60
• The new MAb
pore silica and
unique selectiv
• Fast and high
using MAbPac
• High resolutio
mAb fragments
and ADC mimic
Reference
1.
2.
3.
4.
5.
6.
Valliere-Do
Pan H. et a
Veronese,
McCue JT e
Lu Y. et al. C
Leavy, O. N
List all non-Thermo trademarks
SEQUEST, ActiveX, Eksignet,
Scientific and its subsidiaries.
This information is not intended
property rights of others.
Note • PN21024 ISC 0814S 5
e optimized by either
or adding isopropanol
ase B.
ammonium sulfate concentration to 0.6 M by simply changing
the gradient resulted in a higher resolution separation.
FIGURE 4. Separation of PEGylated mAbs
180
Substances using a New
ation of Proteins
C)
Column
n high-purity,
1
Column:
Format:
Mobile phase A:
150
Mobile phase B:
ca particles
de groups. First, a
f Myoglobin,
motrypsinogen was
hase. A fast 6-minute
ieved using a 4.6 ×
tion (Figure 1).
Absorbance (mAU)
125
Gradient:
100
75
Temperature:
Flow rate:
Inj. volume:
Detection:
Sample:
Peaks:
50
her Scientific, Breda, The Netherlands
ent:
perature:
rate:
olume:
ction:
ple:
s:
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
2 M ammonium sulfate, 100 mM
sodium phosphate, pH 7.0
100 mM sodium phosphate, pH
7.0
Time (min)
-5.0
0.0
1.0
7.0
10.0
%A
100
100
100
0
0
%B
0
0
0
100
100
30 ºC
1.0 mL/min
10 µL
UV (280 nm)
Protein mixture
1) Myoglobin
2) Ribonuclease A
3) Lysozyme
4) α-Chymotrypsinogen A
4
6
8
10
12
14
Retention Time (min)
16
18
%A
40
40
40
0
0
40
40
%B
60
60
60
100
100
60
60
20
300
Column:
Format:
Mobile phase A:
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM
phosphate,
pH 7.0 /
Analysis
of Monoclonal
6 sodium
isopropanol (95:5 v/v)
50 mM sodium phosphate, pH
Mobile phase B:
Gradient:
200
50
Temperature:
Flow rate:
Inj. volume:
Detection:
Sample:
Aggregate
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0 /
isopropanol (95:5 v/v)
50 mM sodium phosphate, pH
7.0 / isopropanol (80:20 v/v)
Time (min)
-5.0
0.0
1.0
15.0
20.0
150
100
%A
100
100
100
0
0
%B
0
0
0
100
100
25 ºC
1.0 mL/min
5 µL (8 mg/mL)
UV (280 nm)
Monoclonal antibody
0
0
2
4
6
8
Retention Time (min)
10
12
14
Analysis of Antibody-Drug Conjugate Mimic
Antibody
and Related drug
Substances
using a New Hydrophobic
Interaction
(HIC) Column
Antibody
conjugates
(ADCs)
are aChromatography
rapidly growing
class
protein therapeutics that target cancer
cells.6
List all non-Thermo trademark
SEQUEST, ActiveX, Eksignet,
Scientific and its subsidiaries.
This information is not intende
property rights of others.
PO21024-EN 0814S
Protein and antibody aggregates are formed either during
product expression in cell culture, downstream processing or
storage. These aggregates may cause undesirable immune
reactions which affect the safety of the drug. SEC is the most
widely used technique for the detection and quantification of
protein aggregates in biological drug products. However, several
researchers have reported the use of HIC for the removal of
protein aggregates.4,5 Figure 5 demonstrates the separation of
monoclonal antibody aggregates from the monomer form on the
MAbPac HIC-10 column. In HIC, aggregates typically elute later
than the main peak due to the increased hydrophobicity.
250
Valliere-Do
Pan H. et a
Veronese,
McCue JT e
Lu Y. et al. C
Leavy, O. N
30 ºC
0.5 mL/min
10 µL (1 mg/mL)
UV (280 nm)
PEGylated mAb
1) Derivative 1
2) Derivative 2
FIGURE 5. Separation of mAb aggregates
mAb
e phase B:
2
Time (min)
-5.0
0.0
1.0
15.0
17.0
17.1
23.0
Analysis of mAb Aggregates
or both
peutics and
on a monoclonal
lution required for
their hydrophilic or
mparison of an intact
10. The MAbPac
Fc fragments and
s. These variant
in these fragments.
mn:
at:
e phase A:
0
Absorbance (mAU)
e phase B:
2
0
otein mixture
mn:
at:
e phase A:
25
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0
50 mM sodium phosphate, pH
7.0
1.
2.
3.
4.
5.
6.
Hydrophobic
of
0
mAb
ase A:
ase B:
:
Time (min)
-5.0
0.0
1.0
15.0
20.0
%A
100
100
100
0
0
%B
0
0
0
100
100
25 ºC
1.0 mL/min
Papain digest:10 µL (1 mg/mL)
Intact mAb:
2 µL (5 mg/mL)
UV (280 nm)
a) Intact mAb
b) Papain digest
ation during
or tryptophan
s of bioactivity of
onitor oxidation to
b products.
oxidized mAb
e A:
e B:
e:
4
6
8
Retention Time (min)
10
12
14
Analysis of Antibody-Drug Conjugate Mimic
Antibody drug conjugates (ADCs) are a rapidly growing class of
protein therapeutics that target cancer cells.6 Hydrophobic
interaction chromatography is often used for the separation of
ADCs since attachment of cytotoxin alters the hydrophobicity of
the antibody. The least hydrophobic unconjugated antibody
elutes first and as the number of attached drugs increases the
elution time of each ADC increases as well. Therefore, HIC is
often used to characterize the distribution of ADC molecules with
different drug-to-antibody ratios (DARs). Figure 6 shows the
separation of a cysteine-conjugated ADC mimic sample on the
MAbPac HIC-10 column. The ADC mimics were conjugates
between a drug mimic and mAb via the sulfhydryl group of
interchain cysteine residues which results in a mixture of drugloaded antibody species with 0 to 8 drugs (Figure 6). The
unmodified mAb and ADCs with DAR values ranging from 2 to 8
are well resolved by the MAbPac HIC-10 column. The best peak
shape and separation were achieved using 5% IPA in mobile
phase A and 20% IPA in mobile phase B at 25ºC
FIGURE 6. Separation of CYS-conjugated ADC mimic
40
3
Column:
Format:
Mobile phase A:
30
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
2 M ammonium sulfate, 100 mM
sodium phosphate, pH 7.0
100 mM sodium phosphate, pH 7.0
Time (min)
-5.0
0.0
1.0
15.0
20.0
%A
60
60
60
0
0
%B
40
40
40
100
100
30 ºC
1.0 mL/min
mAb:
2 µL (2.5 mg/mL)
Oxidized mAb: 4 µL (1.3 mg/mL)
UV (280 nm)
mAb
Oxidized mAb
modern
ases
d reduces
cacy.3 Typically it is
However since the
han one PEG may
ated protein and
of PEGylated
Pac HIC-10
e starting
imply changing
Absorbance (mAU)
ure:
e:
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0 /
isopropanol (95:5 v/v)
50 mM sodium phosphate, pH
7.0 / isopropanol (80:20 v/v)
2
Mobile phase B:
Gradient:
2
20
Temperature:
Flow rate:
Inj. volume:
Detection:
Sample:
Peaks:
10
4
1
5
6
0
0
5
10
15
20
Retention Time (min)
25
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0 /
isopropanol (95:5 v/v)
50 mM sodium phosphate, pH
7.0 / isopropanol (80:20 v/v)
Time (min)
-5.0
0.0
1.0
30.0
35.0
%A
100
100
100
0
0
%B
0
0
0
100
100
25 ºC
0.5 mL/min
5 µL (5 mg/mL)
UV (280 nm)
Cys-conjugated ADC mimic
1) Unconjugated mAb
2) DAR 2
3) DAR 4
4) DAR 6
5) DAR 6
6) DAR 8
30
Conclusion
• The new MAbPac HIC-10 column was developed using wide
pore silica and advanced surface bonding technology to achieve
unique selectivity, high recovery and high efficiency.
• Fast and high resolution separation of proteins was achieved
using MAbPac HIC-10 column.
• High resolution separation of various mAb samples including
mAb fragments, oxidized mAb, PEGylated mAb, mAb aggregates
and ADC mimic were obtained using MAbPac HIC-10.
References
Thermo Scientific Poster Note • PN21024 ISC 0814S 7
1. Valliere-Douglass, J. et al. J.Chromatogr. A (2008) 1214, 81.
6) DAR 8
UV (280 nm)
mAb
Oxidized mAb
0
ase B:
ure:
e:
:
10
15
20
Retention Time (min)
25
30
Conclusion
modern
ases
d reduces
cacy.3 Typically it is
However since the
han one PEG may
ated protein and
of PEGylated
Pac HIC-10
e starting
imply changing
paration.
ase A:
5
MAbPac HIC-10, 5 µm
4.6 㽢100 mm
1.5 M ammonium sulfate, 50
mM sodium phosphate, pH 7.0
50 mM sodium phosphate, pH
7.0
Time (min)
-5.0
0.0
1.0
15.0
17.0
17.1
23.0
%A
40
40
40
0
0
40
40
%B
60
60
60
100
100
60
60
• The new MAbPac HIC-10 column was developed using wide
pore silica and advanced surface bonding technology to achieve
unique selectivity, high recovery and high efficiency.
• Fast and high resolution separation of proteins was achieved
using MAbPac HIC-10 column.
• High resolution separation of various mAb samples including
mAb fragments, oxidized mAb, PEGylated mAb, mAb aggregates
and ADC mimic were obtained using MAbPac HIC-10.
References
1.
2.
3.
4.
5.
6.
Valliere-Douglass, J. et al. J.Chromatogr. A (2008) 1214, 81.
Pan H. et al. Protein Sci. (2009) 18, 424.
Veronese, F.M. and Mero, A. Biodrugs (2008) 22, 315.
McCue JT et al. Bioprocess Biosyst Eng. (2008) 31, 261.
Lu Y. et al. Curr Pharm Biotechnol. (2009)10:427.
Leavy, O. Nat.Rev. Immunol.(2010) 10, 297.
List all non-Thermo trademarks and registered trademarks that appear in the poster. Examples include TMT,
SEQUEST, ActiveX, Eksignet, Mascot. Follow this with: All other trademarks are the property of Thermo Fisher
Scientific and its subsidiaries.
This information is not intended to encourage use of these products in any manners that might infringe the intellectual
property rights of others.
30 ºC
0.5 mL/min
10 µL (1 mg/mL)
UV (280 nm)
PEGylated mAb
1) Derivative 1
2) Derivative 2
PO21024-EN 0814S
www.thermofisher.com
©2016 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its
subsidiaries. This information is presented as an example of the capabilities of Thermo Fisher Scientific products. It is not
intended to encourage use of these products in any manners that might infringe the intellectual property rights of others.
Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local
sales representative for details.
Africa +43 1 333 50 34 0
Australia +61 3 9757 4300
Austria +43 810 282 206
Belgium +32 53 73 42 41
Brazil +55 11 3731 5140
Canada +1 800 530 8447
China 800 810 5118 (free call domestic)
400 650 5118
Denmark +45 70 23 62 60
Europe-Other +43 1 333 50 34 0
Finland +358 9 3291 0200
France +33 1 60 92 48 00
Germany +49 6103 408 1014
India +91 22 6742 9494
Italy +39 02 950 591
Japan +81 6 6885 1213
Korea +82 2 3420 8600
Latin America +1 561 688 8700
Middle East +43 1 333 50 34 0
Netherlands +31 76 579 55 55
New Zealand +64 9 980 6700
Norway +46 8 556 468 00
Russia/CIS +43 1 333 50 34 0
Singapore +65 6289 1190
Sweden +46 8 556 468 00
Switzerland +41 61 716 77 00
Taiwan +886 2 8751 6655
UK/Ireland +44 1442 233555
USA +1 800 532 4752
PN21024-EN 0716S