The following documents are demonstrations of real time applications using this product. You must register to download these.
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.
Substrate specificity screening with UV area imaging detector.pdf
AN004 lab-on-capillary systems
This application note illustrates use of the ActiPix™ D100 for detection of three peptides (angiotensin II, Metenkephalin and Leu-enkephalin) following separation by nanoLC on a 75 m i.d. column. Using a nanoLC cartridge with detection capillary having i.d. 75 m, results are shown for loadings of 0.5 ng each peptide. The S/N for Leu-enkephalin is 63, suggesting that the limit of detection is below 0.05 ng (80 fmol). Detection is also demonstrated in the 20 m i.d. fused silica transfer line, and with loading of 1 ng each peptide on column the S/N for Leu-enkephalin is 4.4. Peak broadening due to Taylor dispersion in the wider bore capillary causes the peaks to be slightly wider than those measured directly in the transfer line.
This application note illustrates on-column detection using a monolithic capillary column 100 μm i.d./ 360 μm o.d. and isocratic elution. Flow resistance in monolithic columns is much lower than in conventional particle packed columns, and a syringe pump is used to drive the flow. For 1 pmol loading of caffeine and 0.5 s time constant, the signal-to-noise ratio is 23. This shows that the ActiPix™ D100 detector has femtomole sensitivity for on-column detection. A particular benefit of imaging through the column bed with the ActiPix™ D100 is that time-displaced averaging used in Paraytec’s proprietary software removes any random fluctuations in the signal due to non-uniformity of the stationary phase and scattering effects. As shown with a sharply fronting peak of caffeine under overload conditions, this occurs without any sacrifice of the 70 μm spatial resolution.
Directions for connecting to Agilent HP3DCE
Presented at PittCon 2007, Chicago.
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
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.
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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