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April 01, 2016

Shrinking protein purification workflows

By Karola Winkel, Product Strategy Manager at GE Healthcare

SDS-PAGE analysis is a popular but time-consuming method for checking sample quality when purifying proteins. Amersham PhastSystem delivers automatically analyzed results from SDS-PAGE gels in one hour. Chromatography system users can benefit from fully automated protein analysis to further enhance efficiency of their protein purification workflows.

Fully automated SDS-PAGE analysis complements automated protein purification

Limitations of manual SDS-PAGE analysis

Sample quality needs to be considered at multiple stages in a protein purification workflow. Whether checking protein purity or measuring protein quantity, analysis steps have to be incorporated into the workflow. SDS-PAGE analysis is a widely used protein purification analysis tool, because it is simple and has a generic ability to separate and detect proteins. But traditional methods require a substantial amount of time and can suffer from high variability between experiments and users.

The benefits of full automation

Automatically analyzed results are delivered from SDS-PAGE gels in just one hour. In addition to shortened time-to-results and less hands-on time, full automation offers the following benefits over manual methods:

  • Broader dynamic range
    Protein samples are prelabeled with Cy5 fluorescent dye, which means that gels do not have to be stained and destained after electrophoresis. Cy5 prelabeling has a broader dynamic range (3−4 orders of magnitude) than standard methods, such as silver, Coomassie™, and SYPRO™ Ruby staining, to detect strong and weak signals simultaneously.
  • Higher sensitivity
    Although it is important to maximize the amount of target protein obtained, the ability to minimize impurities often defines the quality of the sample. The sensitivity of fluorescence detection in Amersham PhastSystem allows quantitation of low protein amounts not detected with other methods such as Coomassie staining. It is possible to reliably quantitate impurity levels down to 0.1% of the target band’s intensity.
  • Higher reproducibility
    The integrated system design combined with quality-controlled gel cards and robust labeling protocols provide results that are highly reproducible. Standardization is further supported by automatic logging of all relevant run parameters during a run.

Support for ÄKTA system users

These benefits mean that Amersham PhastSystem can support your ÄKTA purification system workflows with:

  • Rapid monitoring of protein expression
  • Efficient scouting of chromatography media
  • Accurate optimization of elution conditions
  • Instant determination of molecular weight
  • Reliable and fast analysis of protein purity/impurity levels in just 3 mouse clicks:

Amersham PhastSystem delivers automatically analyzed results from SDS-PAGE gels in just one hour. Fast processing in every step, combined with a built-in laser scanner and automated data analysis, supports quick decision making for efficient protein purification.