Why size and molecular weight characterization is important in protein
Why size and molecular weight characterization is important in protein
Why size and molecular weight characterization is important in protein
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Zeta<strong>size</strong>r<br />
µV<br />
Zeta<strong>size</strong>r<br />
Zeta<strong>size</strong>r<br />
Zeta<strong>size</strong>r Auto Plate Sampler<br />
DLS Plate Sampl<strong>in</strong>g Technology for unparalleled data quality <strong>and</strong> automation<br />
Detailed specifications at www.malvern.com/zeta<strong>size</strong>rAPS<br />
APS<br />
Prote<strong>in</strong> <strong>size</strong>
Zeta<strong>size</strong>r<br />
APS<br />
Designed for prote<strong>in</strong> applications<br />
The Zeta<strong>size</strong>r APS<br />
The Zeta<strong>size</strong>r APS <strong>is</strong> a robust,<br />
simple to operate, Dynamic<br />
Light Scatter<strong>in</strong>g system that<br />
automates measurements of<br />
samples <strong>in</strong> <strong>in</strong>dustry st<strong>and</strong>ard<br />
96- or 384-well plates to add<br />
the power of advanced light<br />
scatter<strong>in</strong>g analys<strong>is</strong> to your<br />
laboratory. A m<strong>in</strong>imum of<br />
user <strong>in</strong>put <strong>is</strong> required to start<br />
the measurements. Improve<br />
accuracy, reliability <strong>and</strong> ease<br />
of use with the most advanced<br />
system available – The Zeta<strong>size</strong>r<br />
APS Auto Plate Sampler<br />
2 The Zeta<strong>size</strong>r APS<br />
Spotlight on prote<strong>in</strong>s<br />
Spotlight on prote<strong>in</strong>s<br />
How do you know if your prote<strong>in</strong> <strong>is</strong> well-behaved <strong>in</strong> certa<strong>in</strong> buffers<br />
or if it aggregates over time <strong>in</strong> others? Is your prote<strong>in</strong> still a monomer<br />
after thaw<strong>in</strong>g or reconstitution after freeze dry<strong>in</strong>g? Are your buffer<br />
conditions favorable to prote<strong>in</strong> crystallization? The Zeta<strong>size</strong>r APS<br />
allows you to <strong>in</strong>vestigate the stability of your prote<strong>in</strong> over a wide<br />
range of developmental conditions with m<strong>in</strong>imal user <strong>in</strong>tervention.<br />
Now you don't have to choose<br />
between quality or automation.<br />
The Zeta<strong>size</strong>r APS offers both.<br />
• Automates your <strong>size</strong> measurements<br />
• Uses only small, recoverable sample<br />
volumes (20�L)<br />
• Automated sample 'sipp<strong>in</strong>g' offers<br />
the ultimate <strong>in</strong> sensitivity for high<br />
quality results<br />
• Uses the world's favorite<br />
user-friendly software for fully<br />
customizable options<br />
• Incorporates dual, <strong>in</strong>dependent<br />
temperature controls for both<br />
plate holder <strong>and</strong> measurement cell<br />
for high accuracy <strong>and</strong> repeatability<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Zeta<strong>size</strong>r<br />
APS<br />
Light Scatter<strong>in</strong>g - more than <strong>size</strong> measurement<br />
The sensitivity, simplicity <strong>and</strong> non-<strong>in</strong>vasive nature of dynamic light scatter<strong>in</strong>g makes<br />
the technique ideally suited for detect<strong>in</strong>g aggregates <strong>and</strong> monitor<strong>in</strong>g prote<strong>in</strong> stability.<br />
Dynamic light scatter<strong>in</strong>g (DLS)<br />
Dynamic light scatter<strong>in</strong>g measures the diffusion<br />
rate of molecules <strong>and</strong> particles <strong>in</strong> solution,<br />
which enables the calculation of hydrodynamic<br />
radius, <strong>size</strong> d<strong>is</strong>tribution <strong>and</strong> homogeneity. The<br />
hydrodynamic radius <strong>is</strong> the <strong>size</strong> of the molecules as<br />
they ex<strong>is</strong>t <strong>in</strong> that particular buffer environment.<br />
The advantage of th<strong>is</strong> technique <strong>is</strong> that the material<br />
can be measured <strong>in</strong> any buffer of your choice. No<br />
sample preparation that can potentially change the<br />
solvation or oligomer<strong>is</strong>ation state <strong>is</strong> required.<br />
The sensitivity of DLS means that changes <strong>in</strong><br />
oligomer<strong>is</strong>ation state are easily detected.<br />
Zeta<strong>size</strong>r APS sampl<strong>in</strong>g schematic<br />
Highly accurate,<br />
temperature-<br />
controlled<br />
flow cell<br />
Detector<br />
3 The Zeta<strong>size</strong>r APS<br />
Sample returned to<br />
well or optionally<br />
rejected to waste<br />
Spotlight on prote<strong>in</strong>s<br />
Laser<br />
Parameters measured <strong>in</strong>clude:<br />
•<br />
•<br />
•<br />
•<br />
Mean hydrodynamic radius, which<br />
<strong>is</strong> sensitive to the presence of even<br />
trace amounts of aggregates<br />
Polyd<strong>is</strong>persity Index <strong>in</strong>dicat<strong>in</strong>g the<br />
presence of a mixture of oligomers<br />
or aggregates<br />
D<strong>is</strong>tribution of <strong>molecular</strong> <strong>size</strong>s<br />
Molecular <strong>weight</strong> <strong>in</strong>formation<br />
estimated from <strong>size</strong> us<strong>in</strong>g<br />
establ<strong>is</strong>hed models<br />
• Melt<strong>in</strong>g po<strong>in</strong>t determ<strong>in</strong>ation<br />
Ultrasonic<br />
level sensor<br />
Sampl<strong>in</strong>g<br />
needle<br />
Temperature-<br />
controlled well<br />
plate holder<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Zeta<strong>size</strong>r<br />
APS<br />
Insight <strong>in</strong>to your prote<strong>in</strong>s<br />
4 The Zeta<strong>size</strong>r APS<br />
Spotlight on prote<strong>in</strong>s<br />
<strong>Why</strong> <strong>size</strong> <strong>and</strong> <strong>molecular</strong> <strong>weight</strong> <strong>characterization</strong><br />
<strong>is</strong> <strong>important</strong> <strong>in</strong> prote<strong>in</strong> research<br />
The real value <strong>in</strong> light scatter<strong>in</strong>g <strong>in</strong>strumentation <strong>is</strong> not the properties that are measured,<br />
but rather how these properties can be used to help solve application problems.<br />
The Zeta<strong>size</strong>r APS can be used dur<strong>in</strong>g bioprocess<strong>in</strong>g to screen for the effects of<br />
environmental factors, such as temperature, ionic strength, prote<strong>in</strong> concentration, the<br />
presence of certa<strong>in</strong> lig<strong>and</strong>s or ions, or pH on a prote<strong>in</strong>’s monomeric or oligomeric state.<br />
In drug target development, the Zeta<strong>size</strong>r APS can help you<br />
screen many buffer conditions, to f<strong>in</strong>d optimal crystallization<br />
conditions <strong>and</strong> speed up the structure determ<strong>in</strong>ation process.<br />
In prote<strong>in</strong> therapeutics, the<br />
presence of aggregates can cause<br />
adverse effects <strong>in</strong> several ways.<br />
Large aggregates are easily<br />
removed by filtration whereas<br />
smaller soluble aggregates are<br />
more elusive. DLS measurements<br />
are exqu<strong>is</strong>itely sensitive to<br />
the detection of these soluble<br />
aggregates due to the fact that<br />
the amount of light scattered<br />
depends so strongly on <strong>size</strong>. Th<strong>is</strong><br />
means that DLS can be used to<br />
ensure successful purification,<br />
crystallization <strong>and</strong> formulation<br />
development.<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Zeta<strong>size</strong>r<br />
APS<br />
Typical applications<br />
Aggregate detection<br />
Light scatter<strong>in</strong>g can be used to detect<br />
aggregates <strong>in</strong> prote<strong>in</strong> solutions, optimize<br />
storage conditions to avoid aggregation,<br />
<strong>and</strong> screen small molecule libraries for<br />
aggregated compounds that could act<br />
as non-specific <strong>in</strong>hibitors. The Zeta<strong>size</strong>r<br />
APS can automate these measurements,<br />
enabl<strong>in</strong>g a higher throughput of samples<br />
<strong>and</strong> free<strong>in</strong>g up time for other activities.<br />
Crystal screen<strong>in</strong>g<br />
Prote<strong>in</strong> purity <strong>is</strong> <strong>important</strong> <strong>in</strong> the production of<br />
crystals for determ<strong>in</strong><strong>in</strong>g the <strong>molecular</strong> structure<br />
by X-ray diffraction. The Zeta<strong>size</strong>r APS requires<br />
only m<strong>in</strong>imal amounts of sample <strong>and</strong> enables<br />
you to automate the screen<strong>in</strong>g of a large number<br />
of buffers until the ideal conditions for prote<strong>in</strong><br />
crystallization are found.<br />
5 The Zeta<strong>size</strong>r APS<br />
Spotlight on prote<strong>in</strong>s<br />
Prote<strong>in</strong> solubility screen<strong>in</strong>g<br />
The Zeta<strong>size</strong>r APS can monitor changes <strong>in</strong> <strong>size</strong>, scatter<strong>in</strong>g <strong>in</strong>tensity<br />
<strong>and</strong> polyd<strong>is</strong>persity over time to optimize conditions for long-term<br />
stability <strong>and</strong> shelf-life.<br />
Identify thermal character<strong>is</strong>tics<br />
Stable <strong>and</strong> accurate temperature control <strong>is</strong> part<br />
of the fundamental design of the Zeta<strong>size</strong>r APS.<br />
St<strong>and</strong>ard procedures facilitate the measurement<br />
of prote<strong>in</strong> <strong>size</strong> <strong>and</strong> scatter<strong>in</strong>g <strong>in</strong>tensity over<br />
a user-def<strong>in</strong>ed range of temperatures. Any<br />
changes <strong>in</strong> prote<strong>in</strong> structure <strong>in</strong> response to<br />
changes <strong>in</strong> temperature will be clearly identified<br />
giv<strong>in</strong>g <strong>in</strong>formation on purity <strong>and</strong> ass<strong>is</strong>t with<br />
studies to improve shelf-life.<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Zeta<strong>size</strong>r<br />
APS<br />
Spotlight on prote<strong>in</strong>s<br />
Software designed for prote<strong>in</strong> special<strong>is</strong>ts<br />
6<br />
Plate Navigator<br />
The plate navigator gives an overview of the data<br />
from the whole plate. Colour cod<strong>in</strong>g allows users<br />
to easily d<strong>is</strong>t<strong>in</strong>gu<strong>is</strong>h between 'hotspots' <strong>and</strong><br />
trends accord<strong>in</strong>g to any selected parameter <strong>and</strong><br />
wells can be labeled with <strong>in</strong>dividual results for<br />
detailed <strong>in</strong>spection. An additional feature enables<br />
the software to <strong>in</strong>clude or exclude any results<br />
accord<strong>in</strong>g to user-def<strong>in</strong>ed limits.<br />
Prote<strong>in</strong> workspace<br />
The default setup of the software <strong>is</strong><br />
customized for the analys<strong>is</strong> of prote<strong>in</strong>s<br />
<strong>and</strong> biomolecules. Everyth<strong>in</strong>g about<br />
th<strong>is</strong> <strong>in</strong>terface, from the parameters<br />
reported, the views of the results,<br />
the units used <strong>and</strong> the measurement<br />
procedures are geared to support the<br />
requirements of the prote<strong>in</strong> special<strong>is</strong>t.<br />
The Zeta<strong>size</strong>r APS<br />
Prote<strong>in</strong> utilities<br />
SOPs<br />
Th<strong>is</strong> set of tools <strong>is</strong> designed to help<br />
the user with experimental design <strong>and</strong><br />
<strong>in</strong>terpretation of the data. Ass<strong>is</strong>tance<br />
<strong>is</strong> available for recommended sample<br />
concentrations <strong>and</strong> ‘what if’ scenarios<br />
are available to provide shape estimates<br />
such as the Perr<strong>in</strong> factor, prolate <strong>and</strong><br />
oblate axial ratios, as well as an estimate<br />
of MW from the <strong>size</strong> measured.<br />
St<strong>and</strong>ard operat<strong>in</strong>g procedures (SOPs)<br />
simplify the way measurements are performed.<br />
Any measurement procedure can be customized<br />
to the specific needs of a test or sample. Each<br />
SOP can be saved so that experiments can be<br />
repeated with identical protocols. The Plate<br />
Scheduler allows multiple SOPs to be assigned<br />
to a s<strong>in</strong>gle plate <strong>in</strong>creas<strong>in</strong>g the potential of<br />
the Zeta<strong>size</strong>r APS to accommodate all your<br />
measurement needs.<br />
Prote<strong>in</strong> wizard<br />
Th<strong>is</strong> wizard <strong>is</strong> an expert report <strong>in</strong> the<br />
software that can assess your sample<br />
from a number of po<strong>in</strong>ts of view. It can<br />
give an idea of the total proportion of<br />
aggregates <strong>in</strong> a sample, the ratios of<br />
possible oligomeric structures, <strong>and</strong> can<br />
also suggest whether a sample might<br />
be suitable for crystallization trials.<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Zeta<strong>size</strong>r<br />
APS<br />
Now you can...<br />
• Automate your <strong>size</strong> measurements us<strong>in</strong>g low sample<br />
volumes while ma<strong>in</strong>ta<strong>in</strong><strong>in</strong>g the ultimate <strong>in</strong> DLS sensitivity<br />
• Simply <strong>in</strong>sert your plate <strong>and</strong> press ‘Start’. Walk-away<br />
automation does the rest<br />
• Use st<strong>and</strong>ard operat<strong>in</strong>g procedure (SOP) based software<br />
to def<strong>in</strong>e the measurement protocol <strong>in</strong> each well or group<br />
of wells<br />
• Analyze small amounts of recoverable sample <strong>and</strong><br />
accurately <strong>and</strong> reproducibly measure the <strong>size</strong> d<strong>is</strong>tributions<br />
of your prote<strong>in</strong>s<br />
• Transfer methods developed <strong>in</strong> a Zeta<strong>size</strong>r batch system<br />
to the APS<br />
...with ease<br />
7 The Zeta<strong>size</strong>r APS<br />
Spotlight on prote<strong>in</strong>s<br />
• Screen for optimal buffer conditions for prote<strong>in</strong><br />
solubility/stability<br />
• Increase your sample throughput<br />
• Control plate <strong>and</strong> sample measurement<br />
temperature separately<br />
• Directly view results <strong>and</strong> graphs with<strong>in</strong> the user<br />
<strong>in</strong>terface of the world’s best sell<strong>in</strong>g, most user-friendly<br />
DLS software<br />
• Customize report pages specific to your needs<br />
• Retrieve your data <strong>and</strong> export it to third-party<br />
analys<strong>is</strong> packages<br />
Improve accuracy, reliability <strong>and</strong> ease of use with the most advanced<br />
system available – The Zeta<strong>size</strong>r Auto Plate Sampler. Designed primarily for<br />
prote<strong>in</strong> special<strong>is</strong>ts, the new Zeta<strong>size</strong>r APS uses Malvern’s unique Dynamic Light<br />
Scatter<strong>in</strong>g (DLS) plate sampl<strong>in</strong>g technologyTM to simplify measurements <strong>and</strong> to<br />
ensure no comprom<strong>is</strong>e <strong>in</strong> sensitivity due to automation.<br />
It’s no co<strong>in</strong>cidence that our user-friendly, flexible <strong>and</strong> <strong>in</strong>tuitive software <strong>is</strong> also the<br />
world’s most popular!<br />
www.malvern.com/zeta<strong>size</strong>rAPS
Specifications<br />
Zeta<strong>size</strong>r APS<br />
Size range (radius) 0.3nm (380Da prote<strong>in</strong>) to 0.5 microns<br />
Sensitivity for <strong>size</strong> measurement 0.1mg/mL of 14kDa prote<strong>in</strong><br />
Molecular <strong>weight</strong> range 380Da to 20MDa<br />
Temperature range 2ºC to 90ºC +/- 0.1ºC<br />
M<strong>in</strong>imum sample volume 20µL<br />
Computer <strong>in</strong>terface USB<br />
Laser 50mW 830nm<br />
Attenuation range 10,000,000:1<br />
Weight 20kg<br />
Dimensions W:D:H, 500mm x 390mm x 320mm<br />
Power 90V-260V AC, 50/60Hz,150W<br />
Dry air 100kPa (1bar), 4mm o.d. tub<strong>in</strong>g<br />
Optional PTC-1 Thermoelectric temperature control unit<br />
Temperature range 4ºC to 40ºC +/- 0.1ºC<br />
Weight 12.2kg<br />
Dimensions W:D:H, 200mm x 385mm x 310mm<br />
Power 100V-240V AC, 50/60Hz, 228W<br />
Ma<strong>in</strong> features<br />
• Compatible with <strong>in</strong>dustry st<strong>and</strong>ard SBS 96- <strong>and</strong> 384-well<br />
microplates. Expensive plates with quartz or glass bottoms<br />
are not required for high quality data<br />
• Simple to operate us<strong>in</strong>g graphical user <strong>in</strong>terface<br />
• Dual temperature control for greatest accuracy<br />
• St<strong>and</strong>ard operat<strong>in</strong>g procedures for repeatability<br />
• Variety of screen <strong>and</strong> pr<strong>in</strong>t data output options<br />
• Expert guidance regard<strong>in</strong>g result quality for added confidence<br />
• Global after sales support network for tra<strong>in</strong><strong>in</strong>g <strong>and</strong> service<br />
• M<strong>in</strong>imal user <strong>in</strong>put required to obta<strong>in</strong> high quality results<br />
Malvern Instruments Limited<br />
Grovewood Road • Malvern<br />
Zeta<strong>size</strong>r<br />
Worcestershire • UK • WR14 1XZ<br />
µV<br />
Telephone: +44 (0)1684 892456<br />
Facsimile: +44 (0)1684 892789<br />
Zeta<strong>size</strong>r<br />
Malvern Instruments Worldwide<br />
Sales <strong>and</strong> service centres <strong>in</strong> over 50 countries.<br />
For details v<strong>is</strong>it www.malvern.com/contact<br />
Zeta<strong>size</strong>r<br />
Malvern Instruments has a policy of cont<strong>in</strong>uous product improvement <strong>and</strong> specifications may change.<br />
Malvern Instruments <strong>is</strong> part of Spectr<strong>is</strong> plc, the Prec<strong>is</strong>ion Instrumentation <strong>and</strong> Controls Company.<br />
Malvern, Zeta<strong>size</strong>r <strong>and</strong> the ‘green hills’ logo are <strong>in</strong>ternational Trade Marks owned by Malvern Instruments Ltd<br />
Detailed specifications at www.malvern.com/zeta<strong>size</strong>rAPS<br />
© 2008 MRK1111-01<br />
APS<br />
Spotlight on prote<strong>in</strong>s<br />
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