As intein technologies become better developed, the potential arises for their application to a wide range of industrial problems. It is now clear that inteins will likely play a significant role in a number of industries, including pharmaceuticals, agriculture, biomaterials, specialty chemicals, and pure research. Indeed, intein-based bioseparation systems already constitute a major advance in the production of purified recombinant proteins for research use. These new purification methods are capable of reliable and efficient performance, and dozens of papers have now been published describing their use with a variety of protein targets at laboratory scale. The development of inteins that can be induced to cleave through changes in pH and temperature has also delivered new options for the design of large-scale purification processes. The power and flexibility of these systems suggest that they might become a core technology in bioseparations at both the research and manufacturing scales.
This chapter examines two significant applications emerging from research and development in the area of intein-facilitated bioseparations. The first is large-scale affinity separations based on IMPACT and other systems described by Chong and Xu in this volume. The scale-up of these methods, although promising, must be considered with regard to the associated practical and economic aspects that will ultimately dictate their success. A second significant potential application of inteins is in proteomics research. In this case, the reliability of intein-based purifications suggests that they might be applied to entire libraries of protein targets in a high-throughput format. Miniaturization is facilitated by the relative simplicity of intein-mediated bioseparations, allowing the development of a high-throughput microfluidic system based on a disposable platform. This device provides a research tool with significant advantages over existing methods for the production of purified protein libraries.
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