Epitope tag and method for detection and/or purification of tagged polypeptides
Provided herein is a novel epitope that can be used as a tag in methods for rapid and effective characterization, purification, and subcellular localization of polypeptides of interest, which comprise the tag. The tag is specifically recognized by an epitope specific antibody, which can be used to detect, capture, quantify, and/or purify polypeptides of interest that are tagged with the epitope. Also provided is novel epitope specific antibody.
1. An isolated epitope peptide consisting of 8 to 25 amino acids, wherein the epitope peptide comprises an amino acid sequence as defined in SEQ ID NO: 1 (RX 4 X 5 AX 7 SX 9 W),
wherein X 4 can be K or a substitution;
wherein X 5 can be A or R or a conservative substitution of A or R;
wherein X 7 can be V or a conservative substitution of V, and
wherein X 9 can be H or a conservative substitution of H.
2. The isolated epitope peptide of claim 1 , wherein the epitope peptide comprises a sequence as defined by SEQ ID NO: 32 (X 1 X 2 RX 4 X 5 AX 7 SX 9 WX 11 X 12 ),
wherein X 1 can be P or A;
wherein X 2 can be D or a conservative substitution of D;
wherein X 4 can be K or a substitution;
wherein X 5 can be A or R or a conservative substitution of A or R;
wherein X 7 can be V or a conservative substitution of V, and
wherein X 9 can be H or a conservative substitution of H; and
wherein X 11 and X 12 can be Q or a conservative substitution of Q.
3. The isolated epitope peptide of claim 1 , having an amino acid sequence as defined by SEQ ID NO: 3 (RKAAVSHW); or having an amino acid sequence as defined by SEQ ID NO:4 (PDRKAAVSHWQQ); or having an amino acid sequence as defined by SEQ ID NO: 5 (PVRSAALSQWSS), or having an amino acid sequence as defined by SEQ ID NO:33 (PDRVRAVSHWSS), or having an amino acid sequence as defined by SEQ ID NO: 34 (ADRVRAVSHWSS).