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Freeze-drying method of neuron-specific enolase

An enolase and specific technology, applied in the field of freeze-drying, can solve the problems of high cost, complex components, unfavorable stability of stabilizers, etc., and achieve the effect of improving preservation effect and detection sensitivity

Active Publication Date: 2021-07-09
SOPHONIX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many types of active ingredients in the stabilizer and the cost is high. In addition, the components used in this invention are complex, and the interaction between different components is not conducive to the stability of the stabilizer to improve the preservation of NSE.

Method used

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  • Freeze-drying method of neuron-specific enolase
  • Freeze-drying method of neuron-specific enolase
  • Freeze-drying method of neuron-specific enolase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A freeze-drying method for neuron-specific enolase, comprising the following steps:

[0040] (1) Prepare buffer

[0041] The buffer solution includes the following components: 16g of Tris, 12g of KCl, 3g of disodium hydrogen phosphate dodecahydrate, 6g of magnesium sulfate, 7g of trehalose, and 15g of BSA.

[0042] The preparation method of described buffer solution, comprises the following steps:

[0043] Weigh Tris, KCl, disodium hydrogen phosphate dodecahydrate, MgSO 4 , trehalose, and bovine serum albumin are added to purified water and stirred until completely dissolved, adjusted to pH 7.0, and fixed to 1000ml; filter with a 0.22μm filter membrane.

[0044] (2) Preparation of freeze-dried product

[0045] Take the NSE recombinant protein and dissolve it in the buffer solution, mix it well and prepare 6 concentrations of NSE reagent as the calibrator, the concentration is 0ng / mL, 1ng / mL, 20ng / mL, 75ng / mL, 150ng / mL, 300ng / mL mL.

[0046] (3) Put the freeze-dried...

Embodiment 2

[0050] A freeze-drying method for neuron-specific enolase, comprising the following steps:

[0051] (1) Prepare buffer

[0052] The buffer solution includes the following components: 4 g of Tris, 25 g of KCl, 1 g of disodium hydrogen phosphate dodecahydrate, 0.1 g of magnesium sulfate, 50 g of trehalose, and 1.0 g of BSA.

[0053] The preparation method of described buffer solution, comprises the following steps:

[0054] (1) Weigh Tris, KCl, disodium hydrogen phosphate dodecahydrate, MgSO 4 , trehalose, and bovine serum albumin are added to purified water and stirred until completely dissolved, adjusted to pH 8.5, and fixed to 1000ml; filter with a 0.22μm filter membrane.

[0055] (2) Preparation of freeze-dried product

[0056]Take the NSE recombinant protein and dissolve it in the buffer solution, mix it well and prepare 6 concentrations of NSE reagent as the calibrator, the concentration is 0ng / mL, 1ng / mL, 20ng / mL, 75ng / mL, 150ng / mL, 300ng / mL mL.

[0057] (3) Put the ...

Embodiment 3

[0061] A freeze-drying method for neuron-specific enolase, comprising the following steps:

[0062] (1) Prepare buffer

[0063] The buffer solution includes the following components: 20 g of Tris, 6 g of KCl, 6 g of disodium hydrogen phosphate dodecahydrate, 10 g of magnesium sulfate, 5.0 g of trehalose, and 50 g of BSA.

[0064] The preparation method of described buffer solution, comprises the following steps:

[0065] (1) Weigh Tris, KCl, disodium hydrogen phosphate dodecahydrate, MgSO 4 , trehalose, and bovine serum albumin are added to purified water and stirred until completely dissolved, adjusted to pH 6.8, and the volume is set to 1000ml; filter with a 0.22μm filter membrane.

[0066] (2) Preparation of freeze-dried product

[0067] Take the NSE recombinant protein and dissolve it in the buffer solution, mix it well and prepare 6 concentrations of NSE reagent as the calibrator, the concentration is 0ng / mL, 1ng / mL, 20ng / mL, 75ng / mL, 150ng / mL, 300ng / mL mL.

[0068] ...

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Abstract

The invention belongs to the technical field of freeze-drying, and particularly relates to a freeze-drying method of neuron-specific enolase. The freeze-drying method comprises the following steps: dissolving NSE in a buffer solution to prepare an NSE reagent, and freeze-drying the NSE reagent in a vacuum freeze-drying machine; wherein the buffer solution is prepared from the following parts: trihydroxymethyl aminomethane, KCl, disodium hydrogen phosphate dodecahydrate, magnesium sulfate, trehalose and BSA (Bovine Serum Albumin). The freeze-drying method can effectively preserve the neuron-specific enolase and is beneficial to keeping the maximum activity of the neuron-specific enolase.

Description

technical field [0001] The invention belongs to the technical field of freeze-drying, and in particular relates to a freeze-drying method of neuron-specific enolase. Background technique [0002] Neuron-specific enolase (Neuron-Specific Enolase, NSE) is an acidic protease with a molecular weight of about 78,000. NSE is one of the important markers of small cell lung cancer, which can be used for differential diagnosis and to detect the therapeutic effect of small cell lung cancer after radiotherapy and chemotherapy. NSE can also be used in the differential diagnosis of neuroblastoma and Wilms tumor, and has high clinical application value in the early diagnosis of neuroblastoma. [0003] NSE is an unstable biologically active substance, which will lose its activity rapidly in the environment of room temperature and 2-8°C. When NSE is used as a kit component, if it is not lyophilized, it will cause detection errors and cause the kit to become invalid. Use a special buffer ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96G01N33/573G01N33/53
CPCC12N9/96C12N9/88G01N33/573G01N33/5306C12Y402/01011G01N2333/988
Inventor 李博飞
Owner SOPHONIX CO LTD
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