Microfluidic In-line Protein Purification

  • Negative stain image of lysed human embryonic kidney cells mixed with apoferritin from equine spleen.

  • Extracted apoferritin linked to a 25 nm iron oxide nanoparticle.

About-+

This project focuses on the development of a protein extraction method for single cells using magnetic nanoparticles. The nanoparticles are coupled to cognitive proteins, e.g., antibodies that bind the protein of interest (i.e., their antigen). A “magnetic trap” retains the nanoparticles in the microfluidic system, allowing the target proteins to be isolated. Further, the magnetic nanoparticles serve as electron dense labels aiding the identification of the target protein (antigen) in larger protein complexes.

More detail+-

Systems biology aims to understand the emergence of biological functions from interaction networks. The stochastic nature of biological systems makes the study of individual cells and their underlying network structure and dynamics indispensable. This requires analytical methods with single cell resolution.

Due to the low concentration of protein components and the lack of amplification techniques, single-cell proteomics still challenges available techniques. Novel and complementary approaches are needed to make advances in the field.

We seek to establish a novel approach for the qualitative and quantitative analysis of the entire proteome of single cells by transmission electron microscopy (TEM). In this framework, we have recently developed a compatible technique to physically lyse individual cells. As the identification of protein complexes and their constituents in the crude cell lysate is difficult, a complementary method to isolate and label specific components of interest is also required. Proof of concept experiments have been performed.