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High-stability micelle type water-soluble quantum dot for coupling of macromolecular proteins

A water-soluble quantum technology with high stability, applied in the fields of nanoscience, bioanalytical chemistry, analytical chemistry, and polymer materials, it can solve problems such as limiting applications and achieve the effect of high colloidal stability.

Inactive Publication Date: 2018-07-27
刘宏飞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the application of water-soluble quantum dots prepared by this method in labeling macromolecular proteins is limited.

Method used

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  • High-stability micelle type water-soluble quantum dot for coupling of macromolecular proteins
  • High-stability micelle type water-soluble quantum dot for coupling of macromolecular proteins
  • High-stability micelle type water-soluble quantum dot for coupling of macromolecular proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation of step 1 oil-soluble quantum dots (CdSe) ZnS:

[0027] According to the literature [J.J.Li, Y.A.Wang, W.Z.Guo, et al.J.Am.Chem.Soc.2003, 125, 12567-12575] to synthesize oil-soluble quantum dots (CdSe) ZnS, the particle size is between 4 and 5nm, The surface is covered with trioctylphosphine oxide, showing strong hydrophobicity.

[0028] Preparation of step 2 triblock amphiphilic polymer:

[0029] Through the step-by-step atom transfer radical polymerization reaction, the amphiphilic polymer polystyrene-polyhydroxyethyl methacrylate-polyethylene glycol methyl ether methacrylate of three blocks is obtained, wherein polystyrene is Hydrophobic block, poly(hydroxyethyl methacrylate) is a cross-linked block containing side chain hydroxyl, polyethylene glycol methyl ether methacrylate is a hydrophilic block, polyethylene glycol methyl ether methacrylate The ends are carboxyl modified. The molecular weight of the three-block amphiphilic polymer is 9500-10500...

Embodiment 2

[0036] The preparation of step 1 oil-soluble quantum dot (ZnSe) CdSe:

[0037] According to the literature [X.H.Zhong, R.G.Xie, Y. Zhang, et al. Chem. Mater. Covered with octylphosphine oxide, it exhibits strong hydrophobicity.

[0038] Preparation of step 2 triblock amphiphilic polymer:

[0039] Through ring-opening polymerization, esterification reaction, click chemistry and reversible addition-fragmentation chain transfer polymerization, a three-block amphiphilic polymer polycaprolactone-polyhydroxypropyl methacrylate-polyethylene glycol is obtained, wherein Polycaprolactone is a hydrophobic block, polyhydroxypropyl methacrylate is a cross-linking block containing a side chain hydroxyl group, polyethylene glycol is a hydrophilic block, and the end of polyethylene glycol is modified with a carboxyl group. The molecular weight of the three-block amphiphilic polymer is 16,000-20,000, and the molecular weight of the hydrophilic cross-linked block polyhydroxyethyl methacrylate...

Embodiment 3

[0045] Example 3: Preparation and purification of highly stable micellar water-soluble quantum dot-macromolecular antibody conjugate

[0046] Human cardiac troponin I (cTnI) monoclonal antibody (anti-cTnI, molecular weight ≈ 150KD) was selected as the model of the coupled macromolecular protein, and the surface carboxyl-terminated high-stable micellar water-soluble antibody prepared in Example 1 was selected. Quantum dots, preparing quantum dot-cTnI monoclonal antibody conjugates that can be used in the diagnosis of heart diseases.

[0047] Prepare the following solution first:

[0048] Preparation of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) solution (0.052M, 10mg / ml): Dissolve 1mg of EDC in 100μL of PBS (0.1 M, pH=7.27).

[0049] Preparation of NHS (N-hydroxysuccinimide) solution (0.087M, 10 mg / ml): Dissolve 1 mg of NHS in 100 μL of PBS (0.1 M, pH=7.27).

[0050] Concrete reaction process is as follows:

[0051]Take 20 μL of surface carboxyl-termi...

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Abstract

The invention relates to a high-stability micelle type water-soluble quantum dot for coupling of macromolecular proteins. The high-stability micelle type water-soluble quantum dot has a typical core-shell structure, a hydrophobic inner core of the high-stability micelle type water-soluble quantum dot is composed of an oil-soluble quantum dot and a hydrophobic block of a tri-block amphiphilic polymer, and a crosslinking layer and a hydrophilic layer of the tri-block amphiphilic polymer are respectively composed of a crosslinking block and a hydrophilic block of the tri-block amphiphilic polymer. Since the crosslinking block contains a hydroxyl group and has hydrophily before crosslinking, crosslinking reaction can be generated in a water-phase environment after micelle formation, and a covalent bond network surrounding the hydrophobic inner core is formed, so that the stability of the high-stability micelle type water-soluble quantum dot is comprehensively increased; meanwhile, the tailend of a hydrophilic segment of the tri-block amphiphilic polymer is terminated by functional groups except the hydroxyl group, and the high-stability micelle type water-soluble quantum dot can be used for coupling of various macromolecular proteins. Since the high-stability micelle type water-soluble quantum dot has extremely high stability, the high-stability micelle type water-soluble quantumdot is particularly suitable for coupling and fluorescent labeling of the macromolecular proteins.

Description

technical field [0001] The invention belongs to the fields of macromolecular materials, analytical chemistry, nanoscience and bioanalytical chemistry, and specifically relates to a high-stability micellar water-soluble quantum dot used for macromolecular protein coupling. Background technique [0002] In recent years, the efficient labeling and imaging of macromolecular proteins has always been a hot spot in clinical disease diagnosis and basic biomedical research. Among the currently existing fluorescent labeling materials, inorganic nanomaterials such as quantum dots have the most excellent fluorescent properties. Quantum dots not only have high fluorescence intensity, narrow and highly symmetrical half-width of emission peaks, larger stoke shifts, and stronger photobleaching resistance, but also the fluorescence emission wavelength of quantum dots can be effectively controlled by material composition and particle size selection. , so that fluorescence with multiple emiss...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/02C09K11/88B82Y40/00B82Y20/00C07K16/18
CPCB82Y20/00B82Y40/00C07K16/18C09K11/025C09K11/88C09K11/883
Inventor 刘宏飞
Owner 刘宏飞
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