We have developed a method for Enzymatic Sortase-assisted Covalent Orientation-specific Restraint Tethering (ESCORT) recombinant proteins onto surfaces directly from cell-lysate. With an improved surface passivation method, we obviate the cumbersome purification steps even for single molecule studies that demand high purity in the sample. We demonstrated high-specificity of the method, high-passivity of the surface and uncompromised functional integrity of anchored proteins using single molecule fluorescence and force-mapping. We anticipate that this method will substantially reduce the investment by way of time, money and energy in the area of single molecule studies.
Srinivasan, S., Hazra, J. P., Singaraju, G. S., Deb, D., & Rakshit, S. (2017). ESCORTing proteins directly from whole cell-lysate for single-molecule studies. Analytical Biochemistry, 535, 35–42. https://doi.org/10.1016/j.ab.2017.07.022