Stimulus-responsive precipitating protein tags have been explored as a method for the non-chromatographic purification of target proteins. In this method, a protein of interest is fused to the tag, expressed, and subsequently exposed to some stimulus. This fusion protein precipitates out of solution, is washed and re-dissolved in solution. Then, if necessary, the protein of interest is cleaved from the tag and the tag is again precipitated, leaving behind purified protein. This approach can be very powerful for rapid bioseparations. However, the most common existing approach using elastic-like peptides requires a stimulus (temperature swings) that is impractical for large scale purification. We will present our work in the development of engineered beta roll-like peptides which reversibility fold and precipitate in the presence of calcium ions. We have used the system to purify maltose binding protein as well as several other important recombinant proteins.

Development of a calcium-responsive beta roll peptide as a purification tag for non-chormatographic recombinant protein purification

243rd National Meeting (2012)

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