SageELF Sample Fractionator for DNA and Protein

This scientific poster provides workflow updates for Sage Science’s SageELF and BluePippin instruments, optimized for full-length transcript sequencing with Iso-Seq.

This application note investigates the use of an off-line preparative electrophoresis device to fractionate Host-cell proteins (HCPs) away from the biotherapeutic protein prior to LC-MS analysis. Host-cell proteins (HCPs) are proteins produced by the host cells during recombinant protein drug production. These HCPs contaminate the biotherapeutic drug protein at low levels (1-100 ppm) even after several steps of extensive purification. Since even low level HCPs can stimulate an immunogenic response, regulatory guidelines mandate that these HCPs must be identified and quantified to protect patient safety. Current HCP mass spectrometry (MS) characterization involves some level of pre-fractionation (i.e., electrophoresis, reversed-phase (RP) separation, 2D RP/RP LC, etc.) prior to analysis.

Although the resolution of LS-MS systems is improving every year, size fractionation of complex samples prior to MS analysis is still very useful for many proteomic studies. Despite the promise of automated 2D LC systems, SDS PAGE remains a popular, but laborious, pre-fractionation method for MS. This poster demonstrates an automated preparative electrophoresis system (the ELF system) to fractionate complex protein samples into 12 contiguous size fractions. The system uses a two-dimensional process: first, proteins are electrophoresed through an SDS agarose gel column to separate the proteins by size, and second, the separated proteins are electrophoresed sideways out of the agarose column into a linear array of membrane-bounded elution modules that are positioned alongside the separation gel column. The fractionated samples are recovered in SDS buffer, and can be processed for LC-MS using commercial detergent removal spin columns and standard trypsin digestion methods.

This application note evaluates how the accuracy of protein quantitation can be affected by fractionation at the protein level. Reproducible quantitation of cellular proteins is an important step in the analysis of cellular physiology. Protein-level fractionation in conjunction with tandem mass tag methods were used, using human gastric carcinoma cell line (MKN-45) as the example.

This application note explains the fractionation of Host-cell proteins (HCPs) away from biotherapeutic protein using an off-line preparative electrophoresis device, allowing for sub-ppm level HCP detection. HCPs are proteins produced by the host cells during recombinant protein drug production and can contaminate the therapeutic drug protein even at very low levels. Detection of HCPs was improved through fractionating both the intact and reduced monoclonal antibody.

Sage Science’s new size selection system, called the ELF (Electrophoretic Lateral Fractionator) is designed to fractionate a single genomic sample into 12 contiguous size fractions. The system uses a two-dimensional process to separate DNA through an agarose column in a first direction, and then move the separated DNA fragments sideways into a linear array of buffer-filled elution modules that are positioned alongside the separation column. The instrument can operate in direct current mode for samples up to mid-single kilobases in size, or in pulsed field mode for samples up to 50 kilobases in size. Fractionation range is controlled by selection of voltage protocol, gel concentration, and run timing. Internal standards can be used to improve run-to-run reproducibility, and the instrument can read fluorescein-labeled markers.

This poster reports a library generation strategy that builds upon the benefits of existing empirical strategies by implementing orthogonal, multidimensional protein and peptide separation techniques to acquire a comprehensive HDMSe spectral library data set for E. coli.

This video describes the new size selection platform from Sage Science. The instrument takes a DNA or protein sample, separates the biomolecules by size, and then automatically fractionate the sample into 12 contiguous sub-fractions.

This is the fifth of a series of instructional videos for users of the SageELF by Sage Science. The SageELF is a DNA or protein fractionator which uses electrophoresis to create up to 12 contiguous fractions from a biological sample.

This the fourth of a series of instructional video for users of the SageELF by Sage Science. The SageELF is a DNA and protein fractionator which uses electrophoresis to create up to 12 contiguous fractions from a biological sample.

This the third of a series of instructional video for users of the SageELF by Sage Science. The SageELF is a DNA and protein fractionator which uses electrophoresis to create up to 12 contiguous fractions from a biological sample.

This is the second of a series of instructional videos for users of the SageELF by Sage Science. The SageELF is a DNA and protein fractionator which uses electrophoresis to create up to 12 contiguous fractions from a biological sample.

This the first of a series of instructional video for users of the SageELF by Sage Science. The SageELF is a DNA or protein fractionator which uses electrophoresis to create up to 12 contiguous fractions from a biological sample.

The latest technologies and methods for the molecular biology lab