Fluorescent reagent for detecting trace gamma-globulin, as well as preparation method and application thereof

A fluorescent reagent and globulin technology, applied in the field of fluorescent biosensors, can solve the problem of low specificity and achieve the effects of high selectivity, enhanced fluorescent signal, and enhanced fluorescent intensity

Inactive Publication Date: 2016-07-20
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Aiming at the problems that existing chemical reagents often require the participation of metal ions in the detection of γ-globulin, and the specificity is not high, and pre-separation is required in the quantitative detection of γ-globulin, one of the purposes of the present invention is to provide a simple A fluorescent reagent that is easy to detect trace amounts of γ-globulin. The fluorescent reagent does not require the participation of metal ions, has a specific “light-up” fluorescent response to γ-globulin, and has high detection sensitivity for γ-globulin. It is very fast and highly selective; the second purpose is to provide a method for preparing a fluorescent reagent for detecting trace amounts of gamma-globulin, which is simple and easy to operate

Method used

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  • Fluorescent reagent for detecting trace gamma-globulin, as well as preparation method and application thereof
  • Fluorescent reagent for detecting trace gamma-globulin, as well as preparation method and application thereof
  • Fluorescent reagent for detecting trace gamma-globulin, as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0046] A method for preparing a fluorescent reagent for detecting trace amounts of serum gamma-globulin, the method steps are as follows:

[0047] (1) TABD-Py-PF 6 preparation of

[0048] Take (1Z,3Z)-1,4-bis(4-methoxycarbonyl)phenyl-1,4-dibromo-1,3-butadiene 100mg, 4-pyridineboronic acid 63.7mg, tetrakis(triphenyl Phosphine) palladium 11.9mg, potassium carbonate 168.9mg, join in the two-necked flask of 250mL, vacuumize, fill with nitrogen three times, then inject 120mL of toluene / methanol volume ratio and be the mixed solvent of 3:1; Under the condition of nitrogen protection, heat React at 77°C for 24 hours, use a mixed solution of dichloromethane and methanol as an eluent, and separate by column chromatography to obtain a yellow solid powder, wherein the volume ratio of dichloromethane and methanol is 20:1. It can be known that the yellow solid powder is TABD-Py by nuclear magnetic resonance spectrometer and mass spectrometer, and its proton nuclear magnetic spectrum and ...

Embodiment 2

[0065] A method for preparing a fluorescent reagent for detecting trace amounts of serum gamma-globulin, the method steps are as follows:

[0066] (1) TABD-Py-PF 6 preparation of

[0067]Take (1Z,3Z)-1,4-bis(4-methoxycarbonyl)phenyl-1,4-dibromo-1,3-butadiene 100mg, 4-pyridineboronic acid 60mg, tetrakis(triphenylphosphine ) palladium 10mg, potassium carbonate 150mg, put into a 250mL two-necked flask, vacuumize and fill with nitrogen three times, then inject 120mL of toluene / methanol with a volume ratio of 2:1 mixed solvent; under nitrogen protection, heat to 70°C After reacting for 18 hours, using a mixed solution of dichloromethane and methanol as an eluent, the yellow solid powder was separated by column chromatography, wherein the volume ratio of dichloromethane and methanol was 20:1. It can be known that the yellow solid powder is TABD-Py by nuclear magnetic resonance spectrometer and mass spectrometer, and its proton nuclear magnetic spectrum and mass spectrometry data a...

Embodiment 3

[0083] A method for preparing a fluorescent reagent for detecting trace amounts of serum gamma-globulin, the method steps are as follows:

[0084] (1) TABD-Py-PF 6 preparation of

[0085] Take (1Z,3Z)-1,4-bis(4-methoxycarbonyl)phenyl-1,4-dibromo-1,3-butadiene 100mg, 4-pyridineboronic acid 70mg, tetrakis(triphenylphosphine ) palladium 20mg, potassium carbonate 200mg, put into a 250mL two-necked flask, vacuumize and fill with nitrogen three times, then inject 120mL of toluene / methanol with a volume ratio of 4:1 mixed solvent; under nitrogen protection, heat to 80°C After reacting for 30 h, using a mixed solution of dichloromethane and methanol as an eluent, a yellow solid powder was obtained by column chromatography, wherein the volume ratio of dichloromethane and methanol was 20:1. It can be known that the yellow solid powder is TABD-Py by nuclear magnetic resonance spectrometer and mass spectrometer, and its proton nuclear magnetic spectrum and mass spectrometry data are as ...

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Abstract

The invention discloses a fluorescent reagent for detecting trace gamma-globulin, as well as a preparation method and application thereof, and belongs to the field of fluorescent bio-sensors. The fluorescent reagent is prepared from TABD-Py-PF6 and a good solvent. The preparation method comprises the following steps: dissolving (1Z,3Z)-1,4-di(4-methoxycarbonyl) phenyl-1,4-dibromo-1,3-butadiene in the good solvent, adding 4-pyridine phenylboronic acid, a catalyst and a basic salt, and reacting to obtain TABD-Py; dissolving the TABD-Py in a solvent, adding iodomethane to synthesize iodate, adding potassium hexafluorophosphate, and reacting to obtain TABD-Py-PF6; dissolving the TABD-Py-PF6 in the good solvent to obtain the fluorescent reagent. The fluorescent reagent does not need participation of metal ions, has a specific lighting-type fluorescent response on gamma-globulin, has high and quick sensitivity on gamma-globulin detection, and has high selectivity. The fluorescent reagent has an excellent visible detection signal, can be completely separated from high-precision instruments according to actual requirement, and can meet current requirement for detecting the gamma-globulin in serum to a great extent.

Description

technical field [0001] The present invention relates to a fluorescent reagent for detecting trace gamma-globulin, a preparation method and its application, in particular to a real-time "light-up" fluorescent reagent and a preparation method for solution fluorescence detection of trace gamma-globulin in serum, belonging to field of fluorescent biosensors. Background technique [0002] Globulin is a common protein that basically exists in all animals. The normal value of globulin is 20-35g / L, and the normal value of γ-globulin is 8.3-23g / L. Globulin has an immune function, so it is also called immunoglobulin. The dynamic changes of protein can reflect the functional state of the liver. After electrophoresis, serum protein can be divided into albumin, α 1 globulin, alpha 2 There are five zones of globulin, β globulin and γ-globulin. γ-globulin is mainly composed of immunoglobulin. Except for immunoglobulin, most of the serum proteins are synthesized by the liver. Diseases w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/06C07D213/55G01N21/64
CPCC07D213/55C09K11/06C09K2211/1029G01N21/6486
Inventor 董宇平刘派王远航陈笛笛董晓燚李汉军佟斌石建兵
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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