FAST AND CHEAP PROTEIN PURIFICATION

Two naturally produced in vivo affinity resin
substitutes have been combined with a self-cleaving
tag to develop two novel affinity-based purification
technologies. The first resin alternative is
polyhydroxybutyrate (PHB) granules and the second is
elastin-like polypeptides (ELP). An engineered
tripartite protein fusion is expressed in E. coli
and purified through simple centrifugation. The
intein self-cleaving tag then releases the target
protein with a mild pH shift.

The two systems have been successfully used at
laboratory scale to purify more than a dozen
proteins varying in size, complexity and activity,
demonstrating the proof of principle, high purity
and yield attainable. Hence, the cost associated
with purification of recombinant proteins is reduced
significantly to effectively that of just the
culture medium. It is expected that this
combination of improved economics and simplicity
will constitute a significant breakthrough in both
large-scale production of purified proteins and
enzymes as well as high-throughput proteomics
studies.

Autorentext

Mahmoud Reza Banki holds a BS in chemical engineering and a BA in applied mathematics from University of California at Berkeley. This work is mostly the outcome of his PhD work at the chemical engineering department at Princeton University and was partially sponsered by a National Science Foundation fellowship.

Klappentext

Two naturally produced in vivo affinity resin substitutes have been combined with a self-cleaving tag to develop two novel affinity-based purification technologies. The first resin alternative is polyhydroxybutyrate (PHB) granules and the second is elastin-like polypeptides (ELP). An engineered tripartite protein fusion is expressed in E. coli and purified through simple centrifugation. The intein self-cleaving tag then releases the target protein with a mild pH shift. The two systems have been successfully used at laboratory scale to purify more than a dozen proteins varying in size, complexity and activity, demonstrating the proof of principle, high purity and yield attainable. Hence, the cost associated with purification of recombinant proteins is reduced significantly to effectively that of just the culture medium. It is expected that this combination of improved economics and simplicity will constitute a significant breakthrough in both large-scale production of purified proteins and enzymes as well as high-throughput proteomics studies.