A novel synthetic epitope tag and its monoclonal antibody for specific and efficient protein detection and purification

Abstract

With a rapid growth of the global protein purification and isolation market, there is a huge demand for developing new ligands and kits to facilitate rapid extraction and purification of target protein samples. Affinity tag is one of the most important tools for protein purification at high quality and purity in both research laboratories and biotechnology industries. Here we introduce a novel 18-residue epitope tag carrying the peptide sequence TKENPRSNQEESYDDNES [MW: 2142.07 g/mol; Isoelectric point: pH(I) 3.84] and its highly specific monoclonal antibody for detection and affinity purification of tagged recombinant proteins. We name this new tag system the “TrackenProbeTM” as inspired from part of its peptide sequence. This synthetic peptide is a unique epitope tag designed to give stringent specificity in detection and minimal structural perturbation to target recombinant proteins being tagged. The sequence of this tag shares no significant conformational and sequence homology to any known protein in nature, thus offering maximum immuno-specificity for antibody recognition. Moreover, the inertness of this peptide tag is unlikely to cause any significant interference with natural protein functions or activities in living cells when incorporated into its target protein. Regarding solubility, computational prediction demonstrated little ordered secondary structure of this highly hydrophilic peptide tag, with optimal solvent accessibility for antibody recognition in aqueous solution. Strong hydrophilicity also enables optimal protein hydration to minimize denaturation of the target recombinant proteins. The high specificity and efficient immuno-labeling make TrackenProbeTM-antibody system an attractive tool for a wide variety of protein detection and purification strategies, e.g. Western blotting, in situ immuno-labeling, affinity chromatography, etc. (Technical details: http://www.versitech.hku.hk/reagents/ne/). We welcome biomedical research and business partnership for advanced solutions in the field.