Dye-labeled protein conjugate and method for preparing the same
AssigneeMatsushita Electric Industrial Co., Ltd.
InventorsNobuyuki Shigetou, Jinsei Miyazaki, Hiroshi Nakayama
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Abstract
The present invention provides a dye-labeled protein conjugate in which a protein conjugate is labeled with a large number of dye molecules. In the dye-labeled protein conjugate, a protein conjugate that includes a protein and an antibody bound thereto via a disulfide bond is labeled with a cyanine dye represented by the formula (1) or the formula (2) given below: where R1 and R2 denote hydrogen or an alkyl group, X denotes a halogen, M denotes hydrogen or an alkali metal, and n represents an integer in a range of 1 to 4.
Claims3
A dye-labeled protein conjugate comprising a protein, an antibody bound to said protein via a disulfide bond to form a protein conjugate, and a cyanine dye represented by the following formula (1) or formula (2):
- where R1 and R2 denote hydrogen or an alkyl group, X denotes a halogen, M denotes hydrogen or an alkali metal, and n represents an integer in a range of 1 to 4,
- said protein conjugate being labeled with said cyanine dye, wherein said cyanine dye is bound to said protein conjugate via a covalent bond of an acyl carbon derived from a succinimidyl group present in said cyanine dye with a nitrogen derived from an amino group present in said protein conjugate.
A method for preparing a dye-labeled protein conjugate, said method comprising the steps of:
- reducing a protein in a neutral or weak alkaline phosphate buffer solution;
- adding an antibody to said buffer solution to prepare a protein conjugate;
- and
- adding a cyanine dye represented by the following formula (1) or formula (2) to said buffer solution to label said protein conjugate with said cyanine dye;
- where R1 and R2 denote hydrogen or an alkyl group, X denotes a halogen, M denotes hydrogen or an alkali metal, and n represents an integer in a range of 1 to 4,
- wherein said cyanine dye is bound to said protein conjugate via a covalent bond of an acyl carbon derived from a succinimidyl group present in said cyanine dye with a nitrogen derived from an amino group present in said protein conjugate.
A method for preparing a dye-labeled protein conjugate, said method comprising the steps of:
- reducing a protein in a neutral or weak alkaline phosphate buffer solution;
- adding a cyanine dye represented by the following formula (1) or the formula (2) to said buffer solution to label the reduced protein with said cyanine dye:
- where R1 and R2 denote hydrogen or an alkyl group, X denotes a halogen, M denotes hydrogen or an alkali metal, and n represents an integer in a range of 1 to 4;
- and
- adding an antibody to said buffer solution to bind said antibody to the reduced protein, wherein said cyanine dye is bound to said protein conjugate via a covalent bond of an acyl carbon derived from a succinimidyl group present in said cyanine dye with a nitrogen derived from an amino group present in said protein conjugate.
Dependencies are inferred from the claim text.