CESI 8000 Plus High Performance Separation-ESI Module• Increased coverage
from limited samples• High-resolution
separation of polar
analytes and criticalPTMs• In-depthcharacterization ofproteoforms andprotein complexes

Expand the reach of your mass spectrometer
The CESI 8000 Plus High Performance
Separation ESI Module

Connecting the CESI 8000 Plus to your MS
is as easy as plug and spray.
Simply click the OptiMS cartridge tip into the source adapter.

The CESI 8000 Plus is a valuable accessory for your mass spectrometer when analyzing charged
and polar compounds in every analytical laboratory. The CESI 8000 Plus:• Consumes minute samples with extraordinary sensitivity• Excels at charged and polar metabolites, peptides and more
• Enables high-resolution separation of proteins and protein complexes• Eliminates ion suppression bias so you can see what you’ve been missing• Is robust and easy-to-use

CESI-MS “showed better separation efficiency
and resolution with 100-fold less sample
consumption compared to an LC-based intact
protein separation.”“CESI-MS is a high-end tool for metabolomics
work in my lab”

John Yates III
The Scripps Research Institute, La Jolla, CA, USA

CESI-MS “shows a separation efficiency forhigh-molecular-mass compounds like larger
peptides and proteins that is remarkably
higher than that seen with any LC method”“...high resolution CZE separation of intact
glycoprotein species coupled to MS hassignificant potential forthe in-depth characterization andquantitative analysis of biopharmaceuticalproteoforms.”

Herbert Lindner
Innsbruck Medical University, Austria

Barry Karger
Barnett Institute, Northeastern University, Boston, MA, USA

Dr. Rawi Ramautar
Leiden Academic Center for Drug Research, The Netherlands

For more information, visit: sciex.com/CESI800plus“With only a 200 fmol injection, CESI-MS/
MS enables high quality mass spectra while
completely characterizing the primarystructure of biotherapeutic proteins in termsof amino acid sequence, glycosylation andpost-translational modifications. This allows
you to see more with less, and strengthens thepower of the methodology in the context ofbiosimilarity assessment.”
Yannis-Nicolas Francois
University of Strasbourg, France“With monoclonal antibodies having becomesuch a significant class of drugs, not just in
the biopharma space but the entire pharma,
capillary electrophoresis has become an
indispensable tool for monitoring productrelated impurities and product variants ofthese blockbuster drugs. Advances like theCESI 8000 now allow for the coupling of CE andMS and provide a solution for the desperately
needed identification of peaks separated bycapillary electrophoresis.”
Timothy Blanc
Eli Lilly

Example of high sensitivity
from a limited sample

Multiple experiments in the time of one,with <1 µL sample injected.

 *)./-0/  )/ ).$/4

3000

800

22278

412

564

1201

1460e51465e51470e5 ($/$*).

146387.58

2500146569.16

2000
1500

 ($/$*).
 ($/$*).
)*3$/$*)

146701.58
146156.71

146719.51

146862.701460e55001465e5146535.62

1000146189.93

500146981.51

0146573.04

1500

146536.36

1000
500

2000

01470e513.50146948.991460e51465e5Mass. Da

1470e51475e5Mass. Da

4050

60

70

80

90

Intact MAb Separation of IgG1 by CESI-MS *)./-0/  )/ ).$/4

950
850

3000
2500
2000
1500
1000
500
0

1460e51465e51470e5Mass. Da

750
650
5503000 ($/$*).

146389.09

146387.582500

2500
146569.16

2000
1500

146156.71

 ($/$*).)*3$/$*)

146719.51

146862.701465e5146535.62

1000146189.93

500146981.51

1460e5146573.04

1500

146701.58

5000

 ($/$*).

146536.36

1000

2000

0

1470e51460e5146948.99

1465e5Mass. Da

39 19

12.50

0

146389.09

650 *)./-0/  )/ ).$/4

49

11.50

500

NanoLC-MS/MSm/z of the peptides (Da)

1500

10.50

1000

2500



50

9.50

1500

CE migration time (min)Concentration Values in µM

02000

Mass. Da30



156 782500

$Ȃ - )/"'4*.4'/$*))(*$!$/$*)+-*!$' .

2500

312

3000

350

625

450

14.50

CESI-MS/MS

Data from multiple samples in single MS detection run.
Kindly provided by Yannis-Nicolas Francois, University of Strasbourg, France.

nanoLC-MS/MS

1475e540

44

Mass. Da

16

20

24

28

32

36

LC elution time (min)nanoLC-MS/MS

1470e5$Ȃ - )/"'4*.4'/$*))(*$!$/$*)+-*!$' .

12

Migration Time (min)For more information, visit: sciex.com/CESI800plus

950

1050

Ion Intensity (%)

The unique multi-segment injection (MSI) feature significantly
increases throughput. Multiple injections are separated by 60
seconds of background electrolyte injections for sequential MSanalysis. This technique is commonly used with MRM targeted
analyte detection.

100

Using only 100 ng of yeast mitochondrial extract, CESI-MS significantly increased
peptide identification, resulting in a substantial increase in the number andvariety of proteins identified.

1100 *)./-0/  )/ ).$/4

CESI-MS is particularly valuable for precious samples such as subcellular fractions, microdialysates,CSF, murine samples, CTCs, needle biopsies and FFPE archives, or highly toxic samples such as ADCs andvenoms.
• Consumes minute samples with extraordinary sensitivity• Unique multi-segment injection increases throughput 10X• Extraordinary sensitivity and high resolution enable maximum information with minimalsample consumption (~50 nL per injection)

CESI-MS/MS

Intact MAb Separation of IgG1 by CESI-MS

1250

m/z of the peptides (Da)More data from less sample
The CESI 8000 Plus High Performance
Separation ESI Module

CESI-MS/MS

Each dot is related to a single MS/MSfragmentation spectrum.

Separate charged / polar molecules and proteins in high resolution

Intact MAb Separation of IgG1 by CESI-MS
Reconstructed Intensity

Measurements of highly charged analytes pose considerable challenges to current technologies. CESI is the method of choice for highresolution separation of polar metabolites, peptides and proteins rich in particular post-translational modifications (such as glycosylation,citrullination, methylation, and phosphorylation).

Electrophoretic separation combined with highly efficient ESI supports a variety ofapplications, from investigating the anionic and cationic metabolome, to the study ofproteoforms, glycoforms, and multimeric protein complexes.
The separation of sugar phosphates in central metabolism and baseline separation ofisobaric positional isomers such as citrate and isocitrate demonstrates CESI-MS’ highresolving power. CESI is also the preferred technology for chiral compound separations
in a cost-effective manner compared to chiral LC columns.

For complex peptide
mixture analysis, CESI
excels at various posttranslational modifications(PTMs) revealing importantbiological functionalvariations.

6E+8CH2OH

21000

H19000HO17000H
O

H
OH

H

H

OH

H
1O

HOO
PO

PO

O

O
H

H
OH

H

H

OH

3E+8

OH

D-Glucose-6-phosphate;pKa (strongest acidic): 1.22

O

D-Glucose-1-phosphate;pKa (strongest acidic): 1.16

15000

O

O

13000

H2C

11000

O

P

O

O
O

9000
H

7000
5000

CH2OH

H

HO

OH

H

OH

D-Fructose-6-phosphate;pKa (strongest acidic): 1.49

3000
1000

12.5

13.0

13.5

14.0

14.5

15.0

15.5

16.0

16.5

17.0

17.5

18.0

Time (Min.)

Kindly provided by Dr. Rawi Ramautar, Leiden Academic Center for Drug Research, TheNetherlands.

For more information, visit: sciex.com/CESI800plus

2000
1500
1000
500
01460e51465e51470e5113

100156161■ Non-modified Peptides (n=688)

134

Deamidations

146389.09

146569.16

2000
1500

Deamidations
146701.58

146156.71

Deamidations
and oxidation

146862.70
146981.51

1460e51465e52000

146573.04

1500146536.36

1000
500

146387.58

2500

2500

00Peptides Identified

Ion Intensity

23000

CH2Ion Intensity

25000

Reconstructed Intensity

3000

O
62500

Mass. Da

Total ion electropherogram

CESI-MS for anionic metabolic profiling

3000

1470e5146535.62

1000
146719.51

5000146189.931460e5Mass. Da

146948.99

1465e51470e51475e5

Mass. Da

■ Phosphorylated Peptides (n=57)

85■ Acetylated Peptides (n=688)

32

9101311Diﬀerent glycosylation and modification profiles

9764432111020130

4076215060

Time (Min.)
Kindly provided by Herbert Lindner, Innsbruck Medical University, Austria.

For deep characterization of proteoforms, glycoforms, charge states and PTMs, CESI front-end technology enables liquid
phase high resolution separation of proteins, metal binding and protein complexes. CESI-MS is an invaluable tool to drive
biopharmaceutical development and biochemical mechanistic studies.

Principles and design of CESICESI technology integrates capillary electrophoresis (CE) withelectrospray ionization (ESI) into a single dynamic process within
the same device.

Schematic of CESI sprayer

Sprayer tip

ESI metal needle

CESI was developed as a new front-end separation and ionization
technique, combining the high efficiency and ultra-low flow of CEwith ESI. This ESI optimization has shown to greatly improve assay
sensitivity while reducing ion suppression bias.1, 2Capillary+The OptiMS cartridge locks into place in the CESI 8000 Plus andthe tip simply clicks into the source adapter, creating an easy-touse plug-and-spray for supported MS brands and models.

–HV

Porous tipSample inlet

Static conductive liquid

4.0E+18

Sensitivity (counts/mol)

3.5E+18
3.0E+18
2.5E+18
2.0E+18
1.5E+18

The CESI 8000 Plus was developed in collaboration with mass spectrometry researchers covering
a number of applications. They sought to expand their range of detection and increase sensitivity.
In order to achieve this, ultra-low flow separation was integrated with mass spectrometry using
proprietary CESI-MS technology.

Zero dead-volume design

The sprayer, located within a protective housing of the OptiMS cartridge, combines an intrinsicallylow-flow CE separation with electrospray ionization (ESI) within a single device.

••••• The CESI OptiMS capillary inlet and outlet are of the same inner diameter

1.0E+18
5.0E+17
0.0E+00~10 nL/min ultra low flow

~50 nL/min low flow

~150 nL/min medium flow~250 nL/min moderate flow~350 nL/min high flow

Infused sample, Angiotensin I at 2 ng/mL in 10% acetic acid. ESI voltage, -1,350 V; detection range, 50-3000 m/z

1. Kelly RT, Page JS, Zhao R, Qian W, Mottaz HM, Tang K, Smith RD, Anal.Chem. 2008, 80, 143-149
2. Schmidt A, Karas M, Dulks T, J., Am. Soc. Mass Spectrom. 2003, 14, 492-500

For more information, visit: sciex.com/CESI800plus• The electrical contact for CE separation as well as for ESI generationis achieved through the ESI metal needle filled with conductive fluid• Ultra low flow at the porous capillary tip instantly generates a fine
ESI spray when a low voltage ~ 1.25 KV is applied• The low voltage of CESI-MS reduces oxidation artifacts

A single continuous separation capillary is contained
within the OptiMS cartridge, with the same internal
diameter from sample to spray. This design:
Does not require lengthy column equilibration
Eliminates sample carry-overReduces sample-to-sample injection time
Increases robustness

Multiple experiments / sample analyses fromas little as 5 μL
The nanoVials have been validated for use with5 μL of sample. With careful pipetting, injections
from as little as 1 µL are possible. CESI-MS has
ultra-low sample consumption, leaving mostof the material available for analysis
by orthogonal techniques.

CESI 8000 Plus mobile design
The CESI 8000 Plus comes equipped with an electric heightadjustable mobile lab bench.
• Switching between nanoLC and CESI is swift and simple

Service, support and consumables
Our customer support organization has access to thelatest product updates, software revisions, methods andrepair procedures. The following is available to further
optimize your use of CESI-MS:• Application training• Operation qualification (OQ)
• Chemistries for MS as well as optical detection

Workflows–assays and method development
Multi-MS connectivity and multi-detection capability
In CESI-MS mode, the CESI 8000 Plus can be connected
to a variety of MS brands and models. In stand-alone CE
mode, UV/VIS, PDA, or LIF detection can be selected.
This allows for the analysis of biologics using our CE-SDS,
cIEF and glycan chemistries.

CE assays and method
development

LIF

UV/VisFor more information, visit: sciex.com/CESI800plus

MS analysis

PDA

 SCIEX Bruker Thermo Waters

CESI 8000 Plus easy-to-use software

Module specifications
Dimensions
Height: 29.2 in (74.2 cm)
Door Open: 38.8 in (98.6 cm)
Width: 25 in (63.5 cm)
Depth: 28.4 in (72.1 cm)
Height adjustable portable lab benchwith memory settings
36 in x 30 in (91.4 cm x 76.2 cm)
Height adjustable from 27 in to 44 in(69 - 112 cm)

Ready for use within minutes

Visual and intuitive

Simply select one of the pre-loaded
methods, follow visuals to placesamples and reagents and hit START.

The layout of the user interface
mimics the instrument to enabledirect tracking of sample contents.

Weight (uncrated)
188 lbs (85.3 kg)
Electrical requirements:
Voltage: 100 - 240 V; 50/60 Hz
Voltage range
1 to 30 kV programmable
at 0.1 kV increments
Current range
3 to 300 μA programmable
at 0.1 μA increments

Flexible yet expandable
Select programmed control or directcontrol to maximize throughput andoptimize separation performance.

For more information, visit: sciex.com/CESI800plus

Operating environment range
15 - 30° C
20 - 60% relative humidity(non-condensing)

Sample trays
2 x 48 CESI vials/0.3 mL microvials
Buffer tray
2 x 36 CESI vials
Sample temperatureadjustment sange4 - 60° C
Capillary temperatureadjustment range
15 - 30° C
Stand-alone capillary temperatureadjustment range
15 - 60° C
Pressure delivery range-5 to +100 psi
Minimum required sample
Volume50 µL when using microvials
5 µL when using nanoVials

Ordering information
A98089: CESI 8000 Plus High Performance Separation ESI Module
Includes CE separation module, OptiMS Sprayer technology andUV detector ready for stand-alone or CESI-MS analysis. Also includescomputer, monitor and system controller pre-loaded with the CESI 8000
Plus software. An electric height adjustable mobile lab bench makes
switching between CESI-MS and other front-end separation instruments
swift and simple.
B07367: Silica Surface OptiMS Cartridge
30 μm ID, 150 μm OD x 90 cm
Contains bare fused silica capillary for high-resolution separationof peptides, glycans, amino acids and nucleotides/nucleosides.
B07368: Neutral OptiMS Cartridge
30 μm ID, 150 μm OD x 90 cm
Contains covalently attached neutral hydrophilic capillary surfaceenabling high resolution separation of intact proteins and protein/
peptide/small molecule/metal complexes.
5043467: nanoVial
5 µL starting volume validated, 1 µL starting volume demonstrated
A59494: LIF Detector Upgrade (optional)
Solid state laser and detector
B68372: PDA Detector Upgrade (optional)
PDA detector
Biologic Characterization Kits

For more information, visit: sciex.com/CESI800plus

390953: SDS-MW Assay Kit/Purity/Sizing
477600: Carbohydrate Labeling and Analysis Assay
A58481: cIEF Peptide Marker Kit (pl Marker Kit)
A80976: Advancied cIEF Starter Kit
CESI OptiMS Adapter Kits
B07363: Adapter for Sciex NanoSpray III Ion Source
B07366: Adapter for Thermo Scientific Nanospray II Ion Source
B83386: Adapter for Thermo Scientific Nanospray Flex and Nanospray
Flex NG Ion Sources
B85397: Adapter for Waters NanoLockSpray and Waters NanoFlow
Ion Sources
B86099: Adapter for Bruker Mass Spectrometers
For more product information on our capillary electrophoresis
systems visit sciex.com/ceproducts
For information on CESI 8000 Plus and supported MS models/sources, contact your SCIEX representative.

The SCIEX clinical diagnostic portfolio is For In Vitro Diagnostic Use. Rx Only. Product(s) not available in all countries.
For information on availability, please contact your local sales representative or refer to https://sciex.com/diagnostics.
All other products are For Research Use Only. Not for use in Diagnostic Procedures. Trademarks and/or registeredtrademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners in the United States and/orcertain other countries. © 2020 DH Tech. Dev. Pte. Ltd. RUO-MKT-03-2678-H 02/2020.
AB SCIEX™ is being used under license.

Headquarters
500 Old Connecticut Path
Framingham, MA 01701 USA
Phone 508-383-7700
sciex.com

International Sales
For our office locations please call the division
headquarters or refer to our website atsciex.com/offices

The Power of Precision