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Streptavidin phycoerythrin freeze stabilizer and its preparation method and application

A streptavidin and phycoerythrin technology, applied in the field of streptavidin phycoerythrin freezing stabilizer and its preparation, can solve problems such as wrong diagnosis results, and achieve simple operation, ingenious design and low cost. Effect

Active Publication Date: 2018-10-26
上海透景诊断科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, it is necessary to provide a streptavidin phycoerythrin freeze stabilizer, which can keep streptavidin phycoerythrin stable under freezing conditions, reduce the attenuation of streptavidin phycoerythrin fluorescence signal, Avoid false diagnostic results in assays such as immunofluorescence

Method used

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  • Streptavidin phycoerythrin freeze stabilizer and its preparation method and application
  • Streptavidin phycoerythrin freeze stabilizer and its preparation method and application
  • Streptavidin phycoerythrin freeze stabilizer and its preparation method and application

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preparation example Construction

[0050] The present invention also provides the preparation method of the above-mentioned streptavidin phycoerythrin freeze stabilizer, in the case where the streptavidin phycoerythrin freeze stabilizer comprises: polyvinylpyrolidone substances and alkaline buffer The preparation method of the streptavidin phycoerythrin freeze stabilizer comprises the steps of: uniformly mixing the polyvinylpyrrolidone and the alkaline buffer.

[0051] In the case where the streptavidin phycoerythrin freezing stabilizer also includes: protein, surfactant, and liquid preservative and / or solid preservative, the streptavidin phycoerythrin freezing stability The preparation method of the agent also includes the step of: mixing the protein, the surfactant, the liquid preservative and / or the solid preservative with the polyvinylpyrrolidone substance and the alkaline buffer uniform. That is to say, the polyvinylpyrrolidone substance, the alkaline buffer solution, the protein, the surfactant, and the ...

Embodiment 1

[0057] Embodiment 1 compares the influence of different alkaline buffers on the stabilizing effect of streptavidin phycoerythrin

[0058] Streptavidin phycoerythrin was added to each group in Table 1, that is, added to different buffers respectively. The buffer composition is shown in Table 1. The final concentration of streptavidin phycoerythrin was 2.5ug / ml. Each group was divided into 4 parts, and after freezing at 0h and 4°C, -20°C, -40°C for 96h respectively, the fluorescence signal intensity was detected with a Luminex 200 flow cytometer.

[0059] Each buffer was prepared as follows:

[0060] 0.01M TE buffer: 10mMTris+1mM EDTA, adjust pH to 8.0 with HCl

[0061] 0.42M boric acid buffer: 0.42M boric acid, adjust pH to 8.0 with NaOH

[0062] 1×PBS buffer solution: 5.3ml 0.2M sodium dihydrogen phosphate solution+94.7ml 0.2M disodium hydrogen phosphate solution

[0063] Table 1 The influence of different buffer solutions on the stabilization effect of SA-PE under differ...

Embodiment 2

[0067] The stabilizing effect of embodiment 2 different substances to SA-PE

[0068] Select different substances, such as PVP (polyvinylpyrrolidones), alcohol amines, trehalose, glycerin, DMSO (dimethyl sulfoxide), sucrose and other substances, and add them to the algae containing 2.5ug / ml streptavidin respectively. In the 0.01M TE buffer solution of red protein (the preparation of 0.01M TE buffer solution is the same as that of Example 1, pH=8.0), and a blank control (containing 2.5ug / ml streptavidin phycoerythrin without adding corresponding substances 0.01M TE buffer) to form stabilizers with different compositions. As shown in Table 2, they were divided into 6 parts, 3 parts were tested directly after preparation (0h), and the average value was taken, and the other 3 parts were tested at After storage at -20°C and -40°C for 96 hours respectively, the fluorescence signal intensity was detected with a Luminex 200 flow cytometer.

[0069] Table 2 Stabilization effect of diff...

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Abstract

The invention provides a streptavidin phycoerythrin freezing stabilizer. The streptavidin phycoerythrin freezing stabilizer comprises a polyvinylpyrrolidone substance and an alkaline buffer solution. Preferably, the content of the polyvinylpyrrolidone substance is 0.1%(v / v)-10%(v / v), and preferably 0.5%(v / v)-1%(v / v). The concentration of the alkaline buffer solution is 0.01M-0.05M. The stabilizer also comprises albumen, a surfactant and a preservative (liquid and / or solid). The invention also provides a preparation method and application of the streptavidin phycoerythrin freezing stabilizer. According to the streptavidin phycoerythrin freezing stabilizer provided by the invention, streptavidin phycoerythrin is kept stable under a refrigeration condition, the attenuation of streptavidin phycoerythrin fluorescence signals is reduced, wrong diagnosis results occurring in a measurement method such as fluorescence immunoassay are avoided, the design is ingenious, the preparation is simple and convenient, the cost is low, and the stabilizer is suitable for large-scale promotion and application.

Description

technical field [0001] The invention relates to the technical field of protein stabilizers, in particular to the technical field of protein freeze stabilizers, in particular to a streptavidin phycoerythrin freeze stabilizer and its preparation method and application. Background technique [0002] Phycoerythrin is a novel fluorescent labeling reagent that is isolated and purified from red algae and is currently commonly used. Under the excitation of specific wavelength, phycobiliprotein can emit strong fluorescence, and its fluorescence intensity is 30-100 times that of fluorescein. It has good light absorption performance and high quantum yield, and has a wide range of excitation and emission in the visible spectral region. [0003] Phycoerythrin is easy to combine with antibodies, biotin, avidin, immune protein and other substances to make fluorescent probes. By detecting the fluorescence emitted by it, it can be used for fluorescence microscopy detection, fluorescence im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/68
CPCG01N33/533G01N33/68
Inventor 李志甲郭安亮姚见儿朱凤左红伟
Owner 上海透景诊断科技有限公司
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