CN105602549A - Fluorescence indicator with long fluorescence lifetime - Google Patents
Fluorescence indicator with long fluorescence lifetime Download PDFInfo
- Publication number
- CN105602549A CN105602549A CN201510981005.1A CN201510981005A CN105602549A CN 105602549 A CN105602549 A CN 105602549A CN 201510981005 A CN201510981005 A CN 201510981005A CN 105602549 A CN105602549 A CN 105602549A
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- China
- Prior art keywords
- fluorescence
- indicator
- fluorescence indicator
- atom
- carboxylate
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Classifications
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
Abstract
The invention relates to a fluorescence indicator with long fluorescence lifetime. Europium is taken as central metal and amino bridged dipyridyl is taken as an auxiliary ligand for the fluorescence indicator. The designed fluorescence indicator can be excited by ultraviolet light, produces high-intensity fluorescence and has large stokes shift which can reach 280 nm; meanwhile, the fluorescence indicator has a time resolution characteristic and can emit fluorescence for a long time after excitation, the fluorescence can reach the microsecond level, and biological background fluorescence of a to-be-tested sample can be eliminated according to the characteristic.
Description
Technical field
The present invention relates to a kind of fluorescence indicator, particularly a kind of fluorescence instruction with high fluorescence lifetimeAgent.
Background technology
Fluorescence indicator refers to a class material that can stimulated emission produces fluorescence, although in the prior artThe material that can produce fluorescence is more, but common fluorescence indicator is subject to the restriction of part and central metal,Its fluorescence lifetime is shorter, generally at nanosecond rank (1ns=10-9S), and, due to part with groupImpact, make its solubility and antagonist can modification scope on be subject to greatly restriction.
Summary of the invention
The deficiency existing for prior art, the object of the present invention is to provide a kind of rare earth coordination thing fluorescenceIndicator, this indicator can be to amino acid, polypeptide, protein, biotin, Avidin, nucleic acid etc.Molecule carries out mark, by LASER Excited Fluorescence indicator, detects the fluorescence intensity of launching, Ke YidingAmount detects the concentration of test substance.
For achieving the above object, the present invention is achieved through the following technical solutions:
A kind of fluorescence indicator, it at least includes following compound:
Wherein, R1For-COOH or-CONH (CH2)nNH2Or-CONH(CH2)nNHCO(CH2)mCOON(COCH2CH2CO) or-CONH(CH2)nNHCOPh-p-NCS；
R2Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R3Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R4Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R5Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R6Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom.
R2～R6The effect of group is mainly the solubility in different solvents and the fluorescence that regulates this compoundLuminous efficiency; R1Effect be: 1) extend the life-span of the fluorescence sending after being excited, can reach micro-Second grade; 2) the bonding convenience of increase fluorescence indicator and antibody (or albumen), to improve fluorescence instructionThe modifiability of agent reduces the bioactive impact of fluorescence indicator antagonist (or albumen) simultaneously; 3)Regulate the solubility of fluorescence indicator in biological reagent.
Preferably, described fluorescence indicator, wherein, n is positive integer.
Preferably, described fluorescence indicator, wherein, m is positive integer.
Preferably, described fluorescence indicator, wherein, described carboxylate is-COOCH3。
Preferably, described fluorescence indicator, wherein, described carboxylic acid is-COOH.
The invention has the beneficial effects as follows: the designed fluorescence indicator of this case can be excited by ultraviolet light, produceRaw fluorescence intensity is high, and stokes displacement is large, can be up to 280nm; This indicator has the time and divides simultaneouslyDistinguish characteristic, after exciting, can send for a long time fluorescence, reach Microsecond grade, can utilize this characteristic to eliminate and treatTest sample biological background fluorescence (biological background fluorescence is held time short after exciting, in nanosecond) originally.
Brief description of the drawings
Fig. 1 is the HPLC testing result figure of embodiment 1.
Fig. 2 is the mass spectrogram of embodiment 2.
Fig. 3 is emission spectrum and the exciting light spectrogram of fluorescence indicator.
Fig. 4 is the fluorescence lifetime resolution chart of fluorescence indicator.
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is described in further detail, to make those skilled in the art's referenceDescription word can be implemented according to this.
Embodiment 1
A kind of fluorescence indicator, it at least includes following compound:
Its synthetic method is as follows:
The HPLC figure of this compound is shown in Fig. 1.
Embodiment 2
A kind of fluorescence indicator, it at least includes following compound:
Its synthetic method is as follows: (taking n=2 as example)
The mass spectrogram of this compound is shown in Fig. 2.
Embodiment 3
A kind of fluorescence indicator, it at least includes following compound:
Its synthetic method is as follows: (with n=2, m=6 is example)
Note: DSS is double amber imide suberate.
Embodiment 4
A kind of fluorescence indicator, it at least includes following compound:
Its synthetic method is as follows: (taking n=2 as example)
Embodiment 5
Antibody labeling
Fluorescence indicator in embodiment 3 or 4 is joined in antibody-solutions to pH approximately 8.0, room temperature shakeSwing 1 hour, use splitter to separate, can obtain the antibody of mark, adopt fluorescence spectrophotometer lightThe examination of degree instrumentation, can detect very strong fluorescence.
Embodiment 6
Biotin labeling
By EDC (1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride) and NHS (N-hydroxylSuccinimide) join in biotin solution and activate 15 minutes, then add the fluorescence in embodiment 2Indicator, shakes 1 hour, uses splitter to separate, and can obtain the biotin of mark. AdoptSepectrophotofluorometer test, can detect very strong fluorescence.
Fig. 3 is fluorescence emission spectrum and the exciting light spectrogram of the arbitrary compound of embodiment 1-4, emission spectrumExcite at 340nm, excitation spectrum detects at 620nm, and test environment is pH=7.0 phosphate buffer.
Fig. 4 is for exciting the fluorescence lifetime resolution chart that in embodiment 1-4, arbitrary compound obtains, at 340nmExcite, detect at 620nm. The life-span of common fluorescent dye is nanosecond rank, and can from Fig. 4Go out, the fluorescent dye life-span of this case be Microsecond grade other.
Although embodiment of the present invention are open as above, it is not restricted to description and enforcement sideListed utilization in formula, it can be applied to various applicable the field of the invention completely, for being familiar with abilityThe personnel in territory, can easily realize other amendment, therefore do not deviate from claim and etc. homotypeEnclose under limited universal, the present invention is not limited to specific details and illustrates here and the figure describingExample.
Claims (5)
1. a fluorescence indicator, it at least includes following compound:
Wherein, R1For-COOH or-CONH (CH2)nNH2Or-CONH(CH2)nNHCO(CH2)mCOON(COCH2CH2CO) or-CONH(CH2)nNHCOPh-p-NCS；
R2Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R3Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R4Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R5Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom;
R6Be selected from the one in H atom, carboxylate, carboxylic acid or halogen atom.
2. fluorescence indicator as claimed in claim 1, is characterized in that, n is positive integer.
3. fluorescence indicator as claimed in claim 1, is characterized in that, m is positive integer.
4. fluorescence indicator as claimed in claim 1, is characterized in that, described carboxylate is-COOCH3。
5. fluorescence indicator as claimed in claim 1, is characterized in that, described carboxylic acid is-COOH.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510981005.1A CN105602549A (en) | 2015-12-24 | 2015-12-24 | Fluorescence indicator with long fluorescence lifetime |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510981005.1A CN105602549A (en) | 2015-12-24 | 2015-12-24 | Fluorescence indicator with long fluorescence lifetime |
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| CN105602549A true CN105602549A (en) | 2016-05-25 |
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| CN201510981005.1A Pending CN105602549A (en) | 2015-12-24 | 2015-12-24 | Fluorescence indicator with long fluorescence lifetime |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106432298A (en) * | 2016-09-05 | 2017-02-22 | 中国科学院苏州生物医学工程技术研究所 | Lanthanide metal ether complex and preparation method and application thereof |
| CN106908432A (en) * | 2017-04-17 | 2017-06-30 | 吉林大学 | A kind of albumen base lysosome fluorescence indicator and its application |
| CN108864095A (en) * | 2018-07-27 | 2018-11-23 | 中国科学院长春应用化学研究所 | A kind of cave-shaped chelating agent and preparation method thereof |
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| US6340747B1 (en) * | 1997-10-03 | 2002-01-22 | Cis Bio International | Fluorescent conjugates of nucleosides or nucleotides, process for their preparation and their uses |
| CN101228441A (en) * | 2005-05-02 | 2008-07-23 | 勒瑞音菲炫股份有限公司 | Spectroscopic method for the detection of analytes |
| US20090162861A1 (en) * | 2006-04-10 | 2009-06-25 | Mathis Gerard | Method for suppressing a fret signal, fret signal suppressor agents and use in a method for multiplexing biological events |
| US20150185150A1 (en) * | 2012-02-22 | 2015-07-02 | Cisbio Bioassays | Method for normalizing the luminescence emitted by a measuring medium |
| CN105218570A (en) * | 2015-07-30 | 2016-01-06 | 中国科学院苏州生物医学工程技术研究所 | A kind of lanthanide series compound and its preparation method and application |
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2015
- 2015-12-24 CN CN201510981005.1A patent/CN105602549A/en active Pending
Patent Citations (6)
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| US5162508A (en) * | 1987-12-18 | 1992-11-10 | Compagnie Oris Industrie | Rare earth cryptates, processes for their preparation, synthesis intermediates and application as fluorescent tracers |
| US6340747B1 (en) * | 1997-10-03 | 2002-01-22 | Cis Bio International | Fluorescent conjugates of nucleosides or nucleotides, process for their preparation and their uses |
| CN101228441A (en) * | 2005-05-02 | 2008-07-23 | 勒瑞音菲炫股份有限公司 | Spectroscopic method for the detection of analytes |
| US20090162861A1 (en) * | 2006-04-10 | 2009-06-25 | Mathis Gerard | Method for suppressing a fret signal, fret signal suppressor agents and use in a method for multiplexing biological events |
| US20150185150A1 (en) * | 2012-02-22 | 2015-07-02 | Cisbio Bioassays | Method for normalizing the luminescence emitted by a measuring medium |
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| DOMINIQUE GUILLAUMONT 等: "Luminescent Eu (III) and Gd (III) Trisbipyridine Cryptates: Experimental and Theoretical Study of the Substituent Effects", 《CHEM PHYS CHEM》 * |
| FRANK SELLRIE 等: "A homogeneous time-resolved fluoroimmunoassay (TR-FIA) using antibody mediated luminescence quenching", 《ANAL. METHODS》 * |
| H. BAZIN 等: "Homogeneous Phosphodiesterase and Hybridization Assays Using Europium Cryptate: Oligonucleotide Conjugates", 《JOURNAL OF FLUORESCENCE》 * |
| JÚLIO G. SANTOS 等: "Theoretical Spectroscopic Study of Europium Tris(bipyridine) Cryptates", 《J. PHYS. CHEM. A》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106432298A (en) * | 2016-09-05 | 2017-02-22 | 中国科学院苏州生物医学工程技术研究所 | Lanthanide metal ether complex and preparation method and application thereof |
| CN106908432A (en) * | 2017-04-17 | 2017-06-30 | 吉林大学 | A kind of albumen base lysosome fluorescence indicator and its application |
| CN106908432B (en) * | 2017-04-17 | 2019-07-26 | 吉林大学 | A kind of albumen base lysosome fluorescence indicator |
| CN108864095A (en) * | 2018-07-27 | 2018-11-23 | 中国科学院长春应用化学研究所 | A kind of cave-shaped chelating agent and preparation method thereof |
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Effective date of registration: 20170908 Address after: Kolding road high tech Zone of Suzhou City, Jiangsu Province, No. 88 215163 Applicant after: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences Applicant after: SHANGHAI KANGPENG TECHNOLOGY CO., LTD. Address before: Suzhou City, Jiangsu province 215163 City kolding Road No. 88 Applicant before: Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences |
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| TA01 | Transfer of patent application right | ||
| CB03 | Change of inventor or designer information |
Inventor after: Bai Pengli Inventor after: Xu Zhi Inventor after: Yin Jian Inventor after: Yin Huancai Inventor after: Chen Mingli Inventor after: Hu Jun Inventor after: Yang Jianhua Inventor after: Sun Zhongyue Inventor after: Zhao Ziqiang Inventor before: Bai Pengli Inventor before: Yin Jian Inventor before: Yin Huancai Inventor before: Chen Mingli Inventor before: Hu Jun |
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Application publication date: 20160525 |
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