Peak Proteins HPLC-FSEC / SEC-MALS Analytical Platform
We are delighted to announce that our new HPLC-FSEC / SEC-MALS platform is now up and running. We’ve chosen an Agilent BioInert HPLC system with a number of detectors, allowing us to monitor proteins by UV absorbance, fluorescence, and light scattering.
We’ll be using the system in two ways – firstly to allow us to do high-resolution SEC with MALS to measure molecular weight, and secondly to enhance our existing FSEC protocols (primarily for membrane proteins).
During our set-up we were able to accurately measure the weight of a protein using only 0.5 µg per injection, and the limit of detection was as low as 30 ng of protein per injection.
Figure 1. Typical SEC-MALS data showing separation of monomer, dimer, and trimer peaks with MALS weight measurements for each
Figure 2. Left: BSA standards demonstrating good agreement of measured protein weight down to 0.15 mg/mL. Right: Series of protein standards from 6.5 to 440 kDa.
Steve Harborne, Principal Scientist and Head of Membrane Protein Science says “Our membrane protein workflow already utilises the powerful technique of FSEC to streamline construct optimisation and detergent selection. This new addition to our toolkit elevates our throughput, enabling us to process an expanded number of samples with overnight automation and direct fluorescence detection whilst also minimising sample consumption. This enhancement improves our workflow efficiency 5-fold, accelerating optimisation and broadening our exploration of experimental parameters.”
Figure 3. FSEC of a fluorescent protein
The BioInert system is compatible with standard protein purification buffers, and the metal-free sample flow path eliminates the worry of proteins interacting with surfaces inside the instrument.
“This expands our current protein analysis package one step further,” says Duncan Smith, Lead Scientist. “We already have a great selection of tools for solving protein problems, including multiple Akta systems for purification, Thermofluor for determining protein melt curves, a fantastic Mass Spectrometry system for protein ID, and the Fida system for measuring protein size in solution. SEC-MALS is complementary to all these techniques, and more than just another QC step it will let us unlock a deeper understanding of the proteins we’re working with – particularly when they’re difficult to handle or behaving unexpectedly!”
Our SEC-MALS platform will be available to existing customers as part of our protein production packages, and also for analysis of samples you’ve produced yourself. If you think SEC-MALS could help with your projects, please get in touch with us at info@peakproteins.com.
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