High Throughput Protein Lab

The High Throughput Protein (HTP) core facility was created to address the challenge of protein production and purification for projects in structural biology and assay development. The primary purpose of the lab is to aid scientists in identifying conditions and constructs for production of recombinant protein in bacterial expression systems by employing high-throughput methods. The use of robotics enables steps of cloning and expression optimization to be carried out quickly and efficiently by eliminating many of the labor-intensive or error-prone steps normally required for these methods.

The High Throughput Protein lab uses ligation-independent cloning (LIC) for rapid and simultaneous creation of up to 96 different constructs encoding a target protein. The ligation and restriction-digestion steps associated with conventional cloning are unnecessary in LIC. Relative to conventional methods, this increases the success rate of cloning and decreases both the experiment time and the cost of reagents.

We optimize expression in 96-well format to quickly determine the expression levels of the constructs created by LIC. We are able to determine which expression conditions produce the highest levels of total or soluble protein by varying parameters such as temperature, cell line, induction conditions, expression vector and growth media.