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.
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.
A comprehensive introduction to the art and science used by the biochemist and process engineer in the design and optimization of large-scale protein-purification processes. Covers the basics of...
This publication details the isolation of proteins from biological materials, techniques for solid-liquid separation, concentration, crystallization, chromatography, scale-up, process monitoring,...
This book provides an in-depth knowledge of protein purification. The book is designed and written to expedite speedy access to important information regarding protein purification and its various...
Proteins are an integral part of molecular and cellular structure and function and are probably the most purified type of biological molecule. In order to elucidate the structure and function of any...
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