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Brochures and application notes
Paraytec Actipix
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Brochures, leaflets & Catalogues
Leaflets
Application Notes
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This application note shows use of the ActiPix™ for determination of hydrodynamic radius of a protein. The method allows proteins to be used in their native form, without any labelling or denaturation. A plug of protein solution is injected into a fused silica capillary, driven through the capillary by application of pressure, and detected using UV area imaging as it passes windows at entrance to and exit from a loop in the capillary. The radius of the protein is determined by analysis of band broadening due to Taylor dispersion. The method is applicable over a wide concentration range and uses only nanolitres of sample.
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This application note describes how the ActiPix™ UV area imaging detector can be used to test biocatalyst substrate specificity towards a mixture of UV active compounds using a continuous engagement electrophoretically mediated microanalysis (EMMA) assay method.
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Substrate specificity screening with UV area imaging detector.pdf
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AN004 lab-on-capillary systems
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Quantum dots are typically fluorescent nanoparticles of high quantum efficiency (see Figure 1). They can be custom synthesized for a wide variety of applications ranging from medical imaging to next generation LCD displays. Paraytec have developed a new approach to determining the hydrodynamic radius of species in solution. This application note is a proof of principle study to demonstrate the use of the ActiPix™ HT Nano-Sizing System for determination of hydrodynamic radii of quantum dot and gold nanoparticle samples provided by the Physics Department at the University of Leeds. Excellent correlation was obtained between experimental and expected values with typical analysis times of less than 10 minutes. This approach is a significantly faster and more cost efficient approach compared to transmission electron microscopy (TEM) which typically takes several hours to perform this analysis. In addition, conventional particle measuring techniques cannot effectively measure down to sub 20 nanometre sizes.
Technical Notes
Latest Technical Notes from Paraytec
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Formula for calculating Taylor Dispersion
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Directions for connecting to Agilent HP3DCE
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Directions for Installing a Capillary Window in a nanoLC Cartridge
Publications
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Our latest article from the June 2008 edition of Chromatography Today
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Chemistry Focus article in International Laboratory, February 2007 discussing how separations in capillaries can be readily integrated with other in-capillary processes such as mixing and reaction, laying the foundations for a versatile and widely applicable lab-on-capillary format
Posters & Presentations
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Pharmacopoeial dissolution methods are the routine test of in-vitro product performance, but whilst they measure product consistency they provide little detailed information about dissolution mechanism. Drug product dissolution is complex, and involves several more fundamental processes such as: hydration, disintegration, erosion, particle de-agglomeration, and dissolution of the drug substance. Thus what is called drug product “dissolution” is actually a combination of several processes. In this poster we present work on the development and application of UV area imaging as a tool to investigate the dissolution of the drug substance. Theories covering the dissolution of chemical substances (e.g. starting from Noyes & Whitney) stress the importance of the concentration gradient between the solid/solution interface and the bulk solution. UV imaging at the micron scale allows this concentration gradient to be investigated.
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The ActiPix™ D100 is a novel analytical instrument using UV area imaging and Taylor dispersion analysis (TDA)1,2 for determining diffusion coefficients and hydrodynamic radii of proteins in solution. The detector monitors broadening of a band of a therapeutic protein or small molecule solution injected into a stream of buffer solution and driven by a syringe pump through a fused-silica capillary. The band is imaged at two windows, the first on entry to and the second on exit from a loop in the capillary. The hydrodynamic radius follows from the measured differences between peak times (first moments) and variances (second moments) at the two windows3. Preliminary experimental work has focused on comparison of this technique with that of dynamic light scattering (DLS) which is routinely used for particle size characterization in solution. Preliminary results indicate good correlation between the two techniques for measuring protein (BSA, Ovalbumin, Lysozyme and IgG) and small molecule (caffeine) hydrodynamic radii. Initial Actipix measurements for protein solutions show hydrodynamic radii results in good correlation with literature values. Repeatability and precision were also tested using model protein and small molecule solutions. The ability to determine protein aggregate levels is being explored.
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In this poster, Paraytec introduce the principles, operation and key benefits of using surface dissolution imaging to the formulations community for rapid drug development. Model compounds are used to demonstrate practicle application of this revolutionary technique.
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Characterization of proteins and their interactions is fundamental to the understanding of biochemical processes since most biomolecular processes involve the molecular recognition and ligand binding by proteins. Protein based drugs represent special challenges in terms of drug development and manufacturing. The structural integrity is sensitive to the conditions encountered during formulation, production and storage. The therapeutic efficacy is closely related to the conformational structure; thus development of protein drugs requires a detailed knowledge of the physical and chemical stability. In this context, it would be beneficial if biophysical and functional characterization could be combined into one method.
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The Bioconversion-Chemistry-Engineering Interface Programme (BiCE) is a multidisciplinary collaboration between three academic Departments at UCL focusing on the integration of biocatalysis and chemistry with engineering. One of the outputs of the BiCE programme are families of evolved enzymes with improved conversion rates and substrate ranges [1]. In parallel we are looking at automated techniques for the high-throughput analysis of these enzymes under process conditions in particular in microwell [2] and microfluidic [3] formats.
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Use of the ActiPix™ UV area imaging detector with multiple windows on a single looped capillary provides new insights with species of pharmaceutical & biopharmaceutical interest. Two key applications are demonstrated: Determination of hydrodynamic radius from imaging pressure driven flow of analyte bands at two windows. Results are shown for proteins and small molecules injected as single species and in mixtures, and for a mixture of proteins with simultaneous electrophoretic separation. Determining substrate specificity of a biocatalyst towards a mixture of UV active compounds using a continuous engagement electrophoretically mediated microanalysis (EMMA) method.
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An A0 poster describing the benifits of the ActiPix™ D100 for determining hydrodynamic radius
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Presented at PittCon 2007, Chicago.
Manuals
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Instructions for Installing and Upgrading the Software for the ActiPix™ D100
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A basic guide to using the ActiPix™ software for CE, LC and Sizing applications
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Information on selecting, installing and maintaining ActiPix™ D100 cartridges
Software
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This zip file contains the latest release of the ActiPix™ Control Software v1.2. We recommend all our existing users upgrade to this version. This download requires a password to open. Please contact Paraytec for access.
What's new
For use in capillary mode:
- Improved lamp stablisation control
- Input/output triggering
- Analogue output channel
- Auto restart for repeat runs
- Rolling trace view
- Frame reprocessing and velocity optimisation
- Ability to compare multiple traces
- Hydrodynamic radius and extinction coefficient calculators
- Improved sizing algorithm
For area imaging mode:
- Image capture with 1x1, 2x2, and 4x4 square pixels (requires Sensor Head upgrade, please contact Paraytec)
- Image data to absorbance conversion
- Image/absorbance profile export functions
- 3D graphics and screen capture
- Diagnostic logging functions
This release contains pre-packaged cartridge and method files for all supported capillaries and applications including Nano-Sizing and Dissolution.
It is recommended that the new cartridge and method files are used for future data collection and processing.
NOTE: This version of the software is 32bit compliant only and will not work with 64bit Windows
For flere detaljer og pris, kontakt venligst MD Scientific via e-mail: info@md-scientific.dk eller telefon: 7027 8565.
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