Labeled protein and method for obtaining the same
It is an object of the present invention to obtain a labeled protein, and specifically, to separate a labeled protein and the same unlabeled protein. There is provided a labeled protein including: a protein to be labeled having a target protein, at least one or more affinity interaction domains for binding to an affinity support, and at least one or more labeling sites; and a labeling reagent binding to at least one of the labeling sites; wherein the affinity of the labeled protein for the affinity support is difference from that of the protein to be labeled for the affinity support.
1. A labeled protein comprising:
a protein to be labeled having a target protein, at least one or more affinity interaction domains for binding to an affinity support, and at least one or more labeling sites; and
a labeling reagent binding to at least one of the labeling sites;
wherein the affinity of the labeled protein for the affinity support differs from that of the protein to be labeled for the affinity support,
wherein the protein to be labeled comprises, as a sequence comprising the at least one or more affinity interaction domains and the at least one or more labeling sites, an amino acid sequence which isselected from the group consisting of SEQ ID NOs: 1 and 4 to 5:
(SEQ ID NO: 1)
His-His-His-His-His-His-Gly-Gly-Cys;
(SEQ ID NO: 4)
Cys-His-His-His-His-His-His-Cys;
and
(SEQ ID NO: 5)
Cys-His-His-His-His-His-His-Gly-Gly-Cys.
wherein the labeling reagent comprises at least one selected from the group consisting of dye, particle, phospholipid and polyethylene glycol-phospholipid complex.
2. A labeled protein comprising:
a protein to be labeled having a target protein, at least one or more affinity interaction domains for binding to an affinity support, and at least one or more labeling sites; and
a labeling reagent binding to at least one of the labeling sites;
wherein the affinity of the labeled protein for the affinity support differs from that of the protein to be labeled for the affinity support,
wherein the protein to be labeled comprises, as a sequence comprising the at least one or more affinity interaction domains and the at least one or more labeling sites, an amino acid sequence which is selected from the group consisting of SEQ ID NOs: 6 to 14:
(SEQ ID NO: 6)
Ala-Ala-Ala-Leu-Glu-His-His-His-His-His-His-Gly-
Gly-Cys;
(SEQ ID NO: 7)
Ala-Ala-Ala-Leu-Glu-His-His-His-His-His-His-Cys;
(SEQ ID NO: 8)
Ala-Ala-Ala-Leu-Glu-Cys-His-His-His-His-His-His;
(SEQ ID NO: 9)
Ala-Ala-Ala-Leu-Glu-Cys-His-His-His-His-His-His-
Cys;
(SEQ ID NO: 10)
Ala-Ala-Ala-Leu-Glu-Cys-His-His-His-His-His-His-
Gly-Gly-Cys;
(SEQ ID NO: 11)
Ala-Ala-Ala-Cys-His-His-His-His-His-His;
(SEQ ID NO: 12)
Ala-Ala-Ala-Gly-Cys-Gly-Gly-Leu-Glu-His-His-His-
His-His-His;
(SEQ ID NO: 13)
Ala-Ala-Ala-Cys-Leu-Glu-His-His-His-His-His-His;
and
(SEQ ID NO: 14)
Ala-Ala-Ala-Leu-Cys-His-His-His-His-His-His.
wherein the labeling reagent comprises at least one selected from the group consisting of dye, particle, phospholipid and polyethylene glycol-phospholipid complex.
3. The labeled protein according to claim 1 , wherein association rate constant K on between the labeled protein and the affinity support is 1/10 or less as compared with association rate constant between the protein to be labeled and the affinity support.
4. The labeled protein according to claim 1 , wherein the target protein is a single-chain antibody.
5. A labeled protein comprising
a protein to be labeled having a target protein, at least one or more affinity interaction domains for binding to an affinity support, and at least one or more labeling sites; and
a labeling reagent binding to at least one of the labeling sites;
wherein the affinity of the labeled protein for the affinity support differs from that of the protein to be labeled for the affinity support,
wherein the target protein is a polypeptide comprising an amino acid sequence of SEQ ID NO: 38:
(SEQ ID NO: 38)
DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS
ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ
GTKVEIKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFN
IKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNT
AYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS.