BR-9462A
Native-condition protein characterization.
For better lead optimization.
PROTEOMELAB XL-A/XL-I
TM
PROTEIN CHARACTERIZATION SYSTEM
ProteomeLab XL-A/XL-I.
By measuring proteins as interacting elements instead of in isolation, the ProteomeLab XL-A/XL-I
accelerates the development of diagnostic markers and therapeutic targets, and delivers a unique system
for protein characterization in solution.
Protein heterogeneity.
The ability to characterize sample heterogeneity is an important part of process development, formulation and QA/QC.
The accompanying graphs are distribution plots with heterogeneity visually indicated by peaks. The position of each
peak indicates the sedimentation coefficient, while the area gives the amount of that species. The graph (left) uses the
g(s)-distribution method to resolve three non-interacting proteins. Levels of <10% by weight can be quantified. For more
in-depth analysis, third party software is available for enhanced application versatility and sensitivity. The graph (right)
is a distribution plot of a highly stressed monoclonal antibody sample, with heterogeneity visually indicated by higher
resolution peaks. Use of the more sophisticated c(s) distribution method provides enhanced resolution by removing
the effects of diffusion. Aggregates or other variants can be quantified at levels well below 1% by weight.
0.05
Main Peak (Monomer) 45.5%
1.0
0.3
0.2
0.1
Heptamer 0.1%
0.4
Hexamer 0.4%
0.5
Pentamer 1.4%
0.00
0.6
Tetramer 5.3%
0.01
0.7
Trimer 14.6%
1600KDa, 10%
0.02
Dimer 30.6%
0.03
0.8
? Free Light Chain 1.4%
? HL Half Molecule 0.8%
100KDa, 50%
g(s*) distribution
0.04
Normalized c(s) [Total Area =1]
800KDa, 40%
0.9
18
20 22
24
0
0
10
20
30
40
Sedimentation Coefficient (Svedbergs)
0
2
4
6
8
10
12
14
16
Sedimentation Coefficient (Svedbergs)
Source: J. S. Philo Ph. D., Alliance Protein Laboratories
A critical step in the proteomics process, characterization
involves putting a protein’s structure and function into context
with all other interactions in a particular biological pathway.
“For monitoring protein aggregation and heterogeneity
in many different projects, my XL-I is invaluable.”
Kurt Drickamer, Ph.D., Department of Biochemistry,
University of Oxford, UK
By analyzing proteins as interacting elements instead of in isolation,
the ProteomeLab XL-A/XL-I more closely approximates true physiological
conditions by considering the protein’s conformation (folded or unfolded),
assembly reversibility (interacting systems), stoichiometry (associative state),
and heterogeneity (aggregation).
An important component of lead optimization is the ability to
evaluate a drug candidate under concentrated “shelf-like”conditions
before it moves into expensive preclinical or clinical trials.
The ProteomeLab XL-A/XL-I functions over a wide range
of concentration, temperature, ionic strength and pH, delivering
more thorough characterization than other methods.
Or
Our unique operating methodology measures the relative change
in the distribution of molecular weights, providing an efficient way
to measure heterogeneity, stoichiometry and self-associating systems.
+
Because the ProteomeLab XL-A/XL-I’s measurements are based
on the first principles of thermodynamics and hydrodynamics,
no standards or calibrations are required. As a result, you spend
less time on setup, and more time on discovery.
To ensure compatibility with the many additional capabilities
and product support provided by third party software, all data
is saved in standard ASCII format.
For easy sample preparation, inject as little as 15µL to120 µL of sample and
reference solutions into opposing positions in the cell assembly, and place the
assembly into the system. Up to 28 different sample/reference pairs can be
studied in a single run, with each having a different set of solute/solvent conditions
(pH, ionic strength, concentration). This provides data for accurate analysis in as
little as three hours. Your sample can be retrieved for additional analysis.
An integrated suite of efficient proteomics solutions.
All of Beckman Coulter’s ProteomeLab offerings are designed to serve as an extension of your thinking,
making it easier for you to do your best science, your way. As a result, you get a complete solution that
puts meaningful, actionable information into your hands, quickly and easily.
Interacting systems.
The ProteomeLab XL-A/XL-I is adept at helping determine yes/no binding of a small molecule to a target protein.
In the top-most example, a small-molecule inhibitor (the green square) of a hormone-receptor interaction could bind to
either the receptor (A) or the hormone (B). The inhibitor uniquely absorbs at 321nm and has a low molecular weight
and therefore does not sediment. If it binds to a protein, however, the bound molecule will sediment with the apparent
molecular weight of the protein at 321nm. The left-hand graph represents sedimentation equilibrium of an inhibitor,
receptor mixture, with the absence of curvature for the inhibitor (red) indicating that the compound does not bind to
the receptor. The right-hand graph shows a pronounced curve, indicating that the inhibitor binds to the hormone.
+
A
+
B
Compound binds to B
Compound does not bind to A
5
0.7
Absorbance, 321nm
Absorbance, 321nm
A
+
B
4
3
A
2
+
1
0
6.4
6.45
6.5
6.55
6.6
MW=59,770 Da
0.6
0.5
B
0.4
0.3
6.4
6.45
6.5
6.55
6.6
Radius, cm
Radius, cm
Source: Michelle Arkin Ph. D., Sunesis Pharmaceuticals, Inc.
Multiple detection systems.
The ProteomeLab XL-A is configured with a scanning UV/Vis detection system, which provides
sensitivity for low-concentration work and selectivity for optimizing detection based on the
sample’s maximum absorbance. The XL-I adds Rayleigh Interference Optics and the capability
to measure the change in refractive index resulting from changes in sample concentration.
This delivers increased accuracy and the ability to examine a greater concentration range with
a wider selection of samples. The XL-I can collect data from both detection systems sequentially,
to provide the broadest spectrum of data for protein characterization in solution.
“There are other bioanalytical instruments and methodologies for
characterizing the behavior of proteins, but the XL-A/XL-I is often my
first choice because of its unique ability to deliver quantitative
information about heterogeneity, conformation and association state.
Without it, we and our clients would waste a lot of time and material
doing calorimetry, SPR, CD, and other procedures on bad samples.”
John Philo, Ph. D., Alliance Protein Laboratories,
Thousand Oaks CA, USA
Molecular conformation.
Cumulative Fraction
Low Salt –Unfolded Structure
1.0
0.8
0.6
0.4
0.2
0.0
20 30 40 50 60
S20,w
Mg2+
Cumulative Fraction
High Salt –Folded Structure
1.0
0.8
0.6
0.4
0.2
0.0
20 30 40 50 60
S20,w
The characterization of molecular
conformation is a powerful quantitative
method for proving to regulatory
agencies that changes in manufacturing
processes produce products with
identical solution conformations.
It’s also used in preclinical studies to
verify that recombinant proteins are
properly folded, and as an aid in
selecting mutants or engineered
forms of a protein.
This example demonstrates molecular
conformation under varying salt
conditions. In TE buffer (top) an
oligonucleosome structure was shown
to have a sedimentation coefficient of
between 27-30 S (visually indicated by
a vertical distribution plot). In 1.8 mM
MgCl2, the same oligonucleosome
structure was shown to have a
sedimentation coefficient of between
33-52 S (visually indicated by a
non-vertical distribution plot).
The solution’s molecular mass of the
oligonucleosome in these two salts’
concentrations remained the same
(demonstrated by sedimentation
equilibrium), indicating that the
oligonucleosome folded into a more
compact conformation in the
presence of salt.
Source: Jeffrey C. Hansen, Ph.D.
Dept. of Biochemistry, University of Texas
Health Science Center, San Antonio,Texas, USA.
The ProteomeLab advantage.
To provide researchers with the most comprehensive and efficient proteomics research solutions, we’re
integrating our leading-edge systems, software and chemistries with a targeted focus on protein research.
By combining our many technological strengths, ProteomeLab delivers a system of complementary tools
that are uniquely optimized for accelerating protein discovery.
Stoichiometry.
Data describing a protein’s stoichiometry can be useful early in drug development, and in providing characterization
and comparability data to regulatory agencies. The data below is for a sequence homolog of a tumor necrosis factor,
part of a family of proteins that are all trimers. Size exclusion chromatography indicated that this protein is a monomer,
suggesting either that it is not a true trimer or that it was not properly folded after expression in E. coli. In this form of
plot, a single species gives a straight line with a slope proportional to the solution mass. The blue line shows the slope
predicted for a monomer, the red for a trimer. Clearly, this protein is indeed a trimer, as expected.
OR
log (absorbance)
0.0
-0.4
-0.8
-1.2
-1.6
-2.0
-0.6
-0.4
-0.2
0.0
0.2
0.4
0.6
(r2 –r02)/2
Source: J. S. Philo Ph. D., Alliance Protein Laboratories
“Yes/No” characterization.
A simple “yes”or “no” answer is not only easy to obtain, many times it’s all that’s
required– especially in initial experiments. While the ProteomeLab XL-A/XL-I
family can accurately measure several decimal places, it also helps save time and
effort by giving you the flexibility to get quick, top-level answers.
“The XL-A was unique in its ability to determine the bivalency of
the Interleukin-6 (IL-6) dimer when binding to the IL-6 receptor
in solution. This information is crucial in interpreting
results from other techniques such as surface plasmon resonance,
which require one of the components to be immobilized.”
Dr. Geoff Howlett, Biochemistry & Molecular Biology
University of Melbourne, Australia
The ProteomeLab XL-A/XL-I is an important part of a broad continuum of Beckman Coulter
products, including automated liquid handling, capillary electrophoresis, centrifugation and
ultracentrifugation, chromatography data systems, DNA sequencing, electrochemistry, HPLC,
integrated core systems, laboratory data management, scintillation counting, and spectrophotometry.
PROTEOMELAB
TM
FROM TISSUES TO TARGETS.
Identify
Software resources –Additional software and algorithms are actively
being developed and are available as shareware to enhance your individual
application needs. The Beckman Coulter website at beckmancoulter.com/xla
provides links to third party software, including: Sedfit–Features the c(s)
algorithm method for measuring heterogeneity at levels below 1% by weight;
Sednterp–A utility program that provides information on amino acids,
common salts and buffers and interpreted results on molecular conformation,
molecular weights and sedimentation coefficients, and; Nonlin–An analysis
program for measuring self-associating or interacting systems for up to
five molecular species.
Isolate
Fractionate
Characterize
Evaluate
Diagnose
Customer training –Our three-day course combines lectures with
hands-on labwork to teach theory, operation and data analysis. You’ll
leave knowing how to efficiently operate and maintain your instrument,
optimize experimental design, save time and minimize sample use.
Service network–As advances in life sciences yield phenomenal improvements, researchers
face growing pressure to do more with less. That’s why Beckman Coulter has maintained
its excellent reputation for service and support. As a result, thousands of our instruments
are installed worldwide. All backed by our on-line, on-site and on-call support.
On-line searchable reference library–See how your colleague is solving a problem.
Explore new methods. Search over 400 references cataloged by author, reference, title
or keyword with our valuable on-line reference resource library.
• Flow cytometry
• Automated fluorescence microscopy
• Microarray technology
• Cell viability analysis
ORDERING INFORMATION
TM
ProteomeLab XL-A/XL-I
Part No.
• General purpose centrifugation
• High performance centrifugation
• Ultracentrifugation
• Centrifugal elutriation
• Automated liquid handling
969340
ProteomeLab XL-I (200-240 VAC, 60 Hz)
includes: An 50Ti (8 hole rotor), 3 assembled cells (sapphire windows),
counterbalance set, torque stand, & acc. kit.
• High performance centrifugation
• Ultracentrifugation
• Liquid chromatography
969341
ProteomeLab XL-I (220-240 VAC, 50 Hz)
includes: An 50 Ti (8 hole rotor), 3 assembled cells (sapphire windows),
counterbalance set, torque stand, & acc. kit.
• Capillary electrophoresis/
Mass spectrometry
• Spectrophotometry
• Protein characterization in solution
• Automated protein crystallization
969342
ProteomeLab XL-A (200-240 VAC, 60 Hz)
includes: An 60 Ti (4 hole rotor), 1 assembled cell (quartz windows),
counterbalance set, torque stand, & acc. kit.
969343
ProteomeLab XL-A (220-240 VAC, 50 Hz)
includes: An 60 Ti (4 hole rotor), 1 assembled cell (quartz windows),
counterbalance set, torque stand, & acc. kit
392773
Individual assembled cell (sapphire windows)
392772
Individual assembled cell (quartz windows)
• Microarray technology
• Software solutions
• Automated immunoassay
• Flow cytometry
• Electrophoresis
• Immunochemistry
• Immunofixation
Specifications
Dimensions
Height: 120.7 cm (47.5 in.)
Width: 94.0 cm (37 in.)
Depth: 67.3 cm (26.5 in)
Weight
465 Kg (1025 lb)
For information on our comprehensive line of ProteomeLab systems, please
contact your local Beckman Coulter representative or visit our web site at:
www.beckmancoulter.com/xla
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Beckman Coulter’s 200,000 installed systems provide essential biomedical intelligence to enhance health care around the world.
Developing innovative solutions in genetic analysis,
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