Tag-Specific Affinity Purification of Recombinant Proteins by Using Molecularly Imprinted Polymers

Analytical Chemistry
Lidia N Gómez-ArribasMaría C Moreno-Bondi

Abstract

Epitope tagging is widely used to fuse a known epitope to proteins for which no affinity receptor is available by using recombinant DNA technology. One example is FLAG epitope (DYKDDDDK), which provides better purity and recoveries than the favorite poly histidine tag. However, purification requires using anti-FLAG antibody resins, the high cost and nonreusability of which restrict widespread use. One cost-effective solution is provided by the use of bioinspired anti-FLAG molecularly imprinted polymers (MIPs). This work describes the development of MIPs, based on the epitope approach, synthesized from the tetrapeptide DYKD as template that affords purification of FLAG-derived recombinant proteins. Polymer was optimized by using a combinatorial approach to select the functional monomer(s) and cross-linker(s), resulting in the best specific affinity toward FLAG and the peptide DYKD. The imprinted resin obtained was used to purify mCherry proteins tagged with either FLAG or DYKD epitopes from crude cell lysates. Both mCherry variants were highly efficiently purified ( R ≥ 95%, RSD ≤ 15%, n = 3) and impurities were removed. Unlike existing antibody-based resins, the proposed tag-imprinting strategy provides a general method for mee...Continue Reading

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Citations

Nov 20, 2019·Journal of Molecular Recognition : JMR·Elena V PiletskaSergey A Piletsky
Feb 6, 2021·Analytical Methods : Advancing Methods and Applications·Huifei ZhongRui Zhao
Apr 23, 2021·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yuanchen LiuYusuke Yamauchi
Nov 8, 2019·Analytical Chemistry·Valérie PichonAudrey Combès
Sep 30, 2020·ACS Applied Materials & Interfaces·Lidia N Gómez-ArribasMaría C Moreno-Bondi

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