A novel peptide tag for detection and purification of recombinant expressed proteins

作者：Jones William T^*; Harvey Dawn; Kirk Christopher; Rakonjac Jasna; Sun Xiaolin; Frearson Nicky; Al Samarrai Taha

来源：Protein Expression and Purification, 2007, 53(2): 404-410.

DOI：10.1016/j.pep.2007.01.006

• peptide tag
• R-tag
• expression vectors
• immunoaffinity purification
• monoclonal antibody
• single chain antibody fragment
• RGL-2
• expression hosts

Peptide tags have proven useful for the detection and purification of recombinant proteins. However cross reactions of antibodies raised to the tag are frequently observed due to the presence of host proteins containing all or parts of the tag. In this report we have identified a unique viral peptide sequence, R-tag, that by blast searches is absent from the commonly expression hosts Arabidopsis thaliana, Escherichia coli, Pichia pastoris and mouse myeloma cell NSO. We have prepared monoclonal antibodies to this peptide and confirmed the absence of this peptide sequence from the above genomes by Western blotting. We have also modified protein expression vectors to incorporate this sequence as a fusion tag in expressed proteins and shown its use to successfully purify recombinant proteins by immunoaffinity procedures.

• 出版日期2007-6