Method for labeling antibody by fluorophore generated by combination of iridium coordination compound and histidine

Abstract

The invention discloses a novel biomolecule labeling method, i.e. taking cyclic iridium coordination compound containing solvent molecules to combine with histidine residue in protein or polypeptide to generate a fluorophore. The method can label polypeptide and bioactive molecules of protein including antibody and the like, and can be further applied to the application field of bioinstrumentation, cell and tissue labeling, biomolecule tracing and the like. Since the fluorescence color of the compound is rich, and the labeling reaction can be completed in normal-temperature and normal butter solution in one step, thus having advantage compared with the commonly-used labeled compound and the labeling method in market, and having wide application prospect.

Description

technical field [0001] The invention relates to a novel biomarking method, that is, through the combination of a kind of self-nonfluorescent metal iridium coordination compound with the histidine residue on the protein to form an excitable fluorescent group, thereby marking the antibody molecule, It is also used in biotechnology fields such as immunoassay detection and immunofluorescence imaging. Background technique [0002] Fluorescent labeling of biomolecules is an important part of applied biotechnology. Biomolecules labeled with fluorescent groups, such as antibodies, polypeptides, and nucleic acids, recognize target molecules through intermolecular interactions, and then use highly sensitive optical methods or electrochemical methods to identify target molecules. Methods (photoluminescence, fluorescence resonance energy transfer, time-resolved, electrochemiluminescence) to obtain signals generated by labeled molecules, in order to form a series of detection methods bas...

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Problems solved by technology

On this basis, by cross-linking fluorescent groups on Ni-NTA, His 6 Sequences with high affinity to Ni-NTA were also developed for His 6 Fluorescent labeling of tagged proteins, but the reported labeling groups have no change in fluorescence intensity before and after binding to the protein. Not only does the sequence need to contain six consecutive histidine residues, but the fluorescent labeling structure is too large, so the application is limited 【4】【5】

The complexation ability of histidine and Cu++ has also been extensively studied. Among them, some researchers used compounds containing conjugated aromatic rings to combine with copper ions to prepare a fluorescent probe, and used the competitive displacement method to detect free histidine in solution. Amino acid, although this method has high sensitivity, it cannot be used to label peptides and proteins in terms of its reaction principle (displacement method). 【6】

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