Leading the Way in Life Science Technologies

GEN Exclusives

More »

Feature Articles

More »
May 1, 2010 (Vol. 30, No. 9)

New Tools Facilitate Protein Expression

Innovative Solutions Seek to Remove Difficult-to-Express Tag from Recombinant Proteins

  • Functional Screening Assays

    Membrane proteins not only can be difficult to express but also to purify and assay. Many researchers approach the study of membrane proteins by over-expressing recombinant fragments of membrane-associated enzymes and studying their activity in solution. However, this can be a daunting challenge for many assay systems, according to Scott Gridley, Ph.D., director of bioproducts at BlueSky Biotech.

    “Membrane-associated proteins derive significant structural, topological, and relational organization from being constrained on the fluid two-dimensional surface of the membrane.”

    The company has developed a technology called TDA 2.0™ that more closely replicates native protein structure to enhance functional assays, Dr. Gridley says. “Template-directed assembly, the process of organizing recombinant protein on a membrane surface, such as a liposome, restores biological context resulting in a more relevant set of data. TDA 2.0 is the only commercially available technology that readily replicates the membrane context in a soluble, fluid, chemically defined system compatible with high-throughput screening.”

    How does it work? According to Dr. Gridley, “to reproduce membrane association, the enzymes are polyhistidine tagged, and the lipid nanospheres are derivatized with nickel-nitrilotriacetic acid so that polyhistidine tagged proteins bind with high-affinity. Membrane proteins in the context of TDA 2.0 naturally form biologically relevant multimers, and interact with untagged interacting partners to form higher-order complexes without any special effort. We have some purified proteins to replicate pathway interactions in the context of TDA 2.0 in a chemically defined system already available.”

    The company currently offers kits for the insulin receptor, insulin-like growth factor receptor tyrosine kinases, and lyn kinase. For the future, Dr. Gridley indicates the company is “developing kits for nearly every receptor tyrosine kinase family member (~70), as well as extending the technology for use with other membrane protein classes.”

    While daunting challenges remain in the arena of difficult-to-express proteins, new paradigms in expression science are helping pave the way for solving those problems.

Related content

Be sure to take the GEN Poll

CRISPR Patent Controversy

Do you think recently released email from a former Broad Institute scientist to Jennifer Doudna will expedite a final legal decision on who owns the CRISPR patent?

More »