Publications [#260984] of Ashutosh Chilkoti

Papers Published

1. Ge, X; Yang, DSC; Trabbic-Carlson, K; Kim, B; Chilkoti, A; Filipe, CDM, Self-cleavable stimulus responsive tags for protein purification without chromatography., Journal of the American Chemical Society, vol. 127 no. 32 (August, 2005), pp. 11228-11229, ISSN 0002-7863 [16089436], [doi]
   (last updated on 2026/01/15)

   Abstract:
   A simple method to purify recombinant proteins is described by fusing a target protein with an intein and an elastin-like polypeptide that only requires NaCl, dithiothreitol, and a syringe filter to isolate the target protein from Escherichia coli lysate. This tripartite fusion system enables rapid isolation of the target protein without the need for affinity chromatography for purification or proteases for cleavage of the target protein from the fusion. The elastin-like polypeptide tag imparts reversible phase transition behavior to the tripartite fusion so that the fusion protein can be selectively aggregated in cell lysate by the addition of NaCl. The aggregates are isolated by microfiltration and resolubilized by reversal of the phase transition in low ionic strength buffer. After resolubilizing the fusion protein, the intein is activated to cleave the target protein from the elastin-intein tag, and the target protein is then isolated from the elastin-intein fusion by an additional phase transition cycle.