MStern blotting: An improved method of sample preparation for mass spectrometry-based proteomics
Inventors: Hanno Steen, Sebastian Berger, Saima Ahmed
Invention Types: Research Tool
Keywords: Assay, ProteomicsFor More Information Contact: Caron, Connie
The ability of mass spectrometry-based proteomics to identify and quantify large numbers of proteins from diverse samples has increasing value in both research and clinical settings. Developed in 2009, Filter Aided Sample Preparation (FASP), which utilizes ultrafiltration membranes, is the current accepted standard for the processing of samples for proteomic analysis.
Researchers in Dr. Hanno Steen’s laboratory at Boston Children’s have developed a novel sample processing workflow, MStern blotting, which retains many of the strengths of FASP while overcoming several of its major limitations. MStern blotting relies on the physical adsorption of proteins onto a membrane instead of size exclusion due to ultrafiltration membranes. This reduces the force required for efficient liquid transfer, and decreases the processing time for 96-well plates, a common high-throughput format, such that 96 samples can be completely processed within less than a workday. The MStern method has been tested on a variety of samples (urine, cerebrospinal fluid, and whole cell lysates) and has been demonstrated to produce comparable or even better results as compared to FASP.
• High-throughput, easily automated sample preparation for proteomic analysis of a wide array of samples
• MStern blotting uses membranes with larger pore sizes than FASP, which allows for efficient liquid transfer for 96-well plates through a vacuum manifold and eliminates prolonged centrifugation steps
• Method removes the need for reverse phase-based desalting of digests prior to LC/MS analysis by elution though organic solvents
Key Publications: Berger ST, Ahmed S, Muntel J, Cuevas Polo N, Bachur R, Kentsis A, Steen J,
Steen H. MStern Blotting-High Throughput Polyvinylidene Fluoride (PVDF)
Membrane-Based Proteomic Sample Preparation for 96-Well Plates. Mol Cell
Proteomics. 2015 Oct;14(10):2814-23.
IPStatus: Pat. Pend.