Summary
This is a new tag system which is suited to protein purification. The characteristics of being capable of eluting the tag peptide from the affinity column (immobilized antibody) using a mild organic solvent enables not only inexpensive protein purification without damaging the protein but also recycling of the affinity column.
We conducted an experiment to purify recombinant protein (band indicated by the arrow in the figure below) which is secreted into the culture medium from transiently transfected HEK293T cells. Lanes 1 to 7 in the figure show the results of cases in which Target tag was used, and Lane 8 shows the result when His tag was used for comparison. Protein purified by the Target tag contained much less contaminating proteins than that purified using His tag. In addition, complete elution of the protein from the affinity column was achieved using propylene glycol, which is a mild organic solvent, in Lane 6.
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Advantages
We established a mouse IgG monoclonal antibody P20.1 which recognizes a stretch of peptide containing the sequence motif YPGQ. The tag peptide (called Target tag) has a sequence with several repeats of YPGQ. Since the binding between the Target tag and P20.1 antibody can be disrupted by a
mild organic solvent, Target tag-based protein purification is not only mild (i.e., do not damage the protein) and inexpensive, but also makes it easier to recycle the affinity column.
Inventors
Prof. Junichi TAKAGI, et al., Osaka University
Patent Status
Issued: JP 5257997, US 8975384, EP 2239267(GB, DE, FR)
Proposal
Available for non-exclusive licensing.
Non-exclusive evaluation for defined period (set up for options).
Collaborative/supportive research.
Product No: TP0036