RT Journal Article
T1 Tag-specific affinity purification of recombinant proteins by using molecularly imprinted polymers
A1 Gómez-Arribas, Lidia
A1 Urraca Ruiz, Javier
A1 Benito Peña, María Elena
A1 Moreno Bondi, María Cruz
AB 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 polyhistidine tag. However, purification requires using anti-FLAG antibody resins, the high cost and non-reusability 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 meeting the growing demand for efficient, inexpensive, and versatile materials for tagged proteins purification.
PB American Chemical Society
SN 0003-2700
YR 2019
FD 2019
LK https://hdl.handle.net/20.500.14352/93327
UL https://hdl.handle.net/20.500.14352/93327
LA eng
NO Lidia N. Gómez-Arribas, Javier L. Urraca, Elena Benito-Peña, and María C. Moreno-BondiAnalytical Chemistry 2019 91 (6), 4100-4106DOI: 10.1021/acs.analchem.8b05731
NO Ministerio de Economía y Competitividad (España)
DS Docta Complutense
RD 7 abr 2025