Water-soluble fluorene compound and application thereof in protein analysis

A compound and protein technology, applied in the field of biological detection and protein detection, to achieve fast speed, improve detection quality and efficiency, and strong anti-interference effect

Active Publication Date: 2017-06-30
西安百萤生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fluorene compounds are difficult to dissolve in water in neutral or slightly acidic environments, and cannot be directly used for the detection of protein substances

Method used

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  • Water-soluble fluorene compound and application thereof in protein analysis
  • Water-soluble fluorene compound and application thereof in protein analysis
  • Water-soluble fluorene compound and application thereof in protein analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1. Synthesis of water-soluble fluorene compound

[0035] The first step: synthetic structural formula (II) compound

[0036] Synthesize following structural formula (II) compound with reference to U.S.Pat 8471035 method

[0037]

[0038] Second step: synthetic structural formula (III) compound

[0039]

[0040] Add 6.3 g of the compound of structural formula (II) obtained in the previous step, 0.32 g of p-toluenesulfonic acid, 20 mL of ethylene glycol and 100 mL of toluene into a 500 mL round bottom flask. Heat to reflux for 8 hours, cool to room temperature, wash the reaction mixture with saturated sodium bicarbonate solution and saturated brine successively, dry the organic phase with anhydrous sodium sulfate, and filter. The filtrate was transferred to a 500mL round bottom flask, 0.74g of TBAB was added, the reaction system was cooled to 0°C under a nitrogen atmosphere, and 9mL of tert-butyl acrylate and 24mL of 50% sodium hydroxide solution were a...

Embodiment 2

[0046] Embodiment 2: Two kinds of transformed forms of structural formula (I ') compound

[0047] 1. When there are ammonia ions in the solution to be detected, the fluorene compound (I') has the following dynamic transformation, and there is an ammonium salt form, that is, M is an amino ion

[0048]

[0049] 2. When there are metal ions in the solution to be detected, the fluorene compound (I') has the following dynamic transformation, and there is a metal ion salt form, that is, M is a metal ion

[0050]

Embodiment 3 Embodiment 1

[0051] Embodiment 3. detect bovine serum albumin with embodiment 1 compound

[0052] 1. Method

[0053] 1. Prepare a series of samples containing serum albumin at different concentrations with protein contents of 10ng / ml, 20ng / ml, 50ng / ml, 100ng / ml, 200ng / ml and 500ng / ml.

[0054] 2. Add the fluorene compound (compound of structural formula (I')) prepared in Example 1 to each sample, so that the final concentration of the fluorene compound in each sample is 1 μM.

[0055] 3. Incubate all serum albumin samples containing the same concentration of fluorene compound at 37°C for 1 hour.

[0056] 4. Measure the fluorescence intensity of these samples containing serum albumin with different concentrations with a spectrophotometer, the samples are excited at 480nm, and the fluorescence is detected at 620nm.

[0057] 2. Results

[0058] The result is shown in Figure 4. The results showed that the fluorescence excitation intensity was high, and the detection results presented a sta...

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Abstract

The invention discloses a water-soluble fluorene compound and application thereof in protein analysis, and belongs to the technical field of biological detection analysis. The water-soluble fluorene compound is a compound as shown in a structural formula I. The water-soluble fluorene compound has the advantages that the fluorescence of the water-soluble fluorene compound having extremely weak fluorescence in a hydrate solution is greatly reinforced after being combined with protein, and the intense fluorescence variation characteristic can be used for quickly detecting protein and analyzing the content of protein. The compound has strong interference resistance, and can be used for effectively eliminating the influence of common non-protein interfering substances in a detection system on a detection result.

Description

technical field [0001] The invention relates to a water-soluble fluorene compound and its application in protein analysis, belonging to the technical fields of biological detection and protein detection. Background technique [0002] Fluorescence-based biomarker detection methods have the advantages of high specificity, high sensitivity, high accuracy, and wide dynamic range, and are widely used in biology, chemistry, medicine, environmental protection and other fields. There are two types of methods for the determination of proteins using fluorescence techniques. One is to measure the autofluorescence of proteins, which is quenched by adding substances; the other is to use fluorescent indicators, that is, fluorescent probes. In order to obtain various information about proteins, proteins are often interacted with fluorescent probes, mostly some fluorescent dyes, and sometimes other reagents are used. After combining with proteins, the spectrum presents energy transfer chara...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C309/15C09K11/06G01N21/64C12Q1/37C12Q1/56
CPCC07C309/15C09K11/06C09K2211/1011C12Q1/37C12Q1/56G01N21/6402G01N21/6486
Inventor 彭若谷伍谊波第五振军
Owner 西安百萤生物科技有限公司
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