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A bovine serum albumin detection probe and its preparation method and application

A technology for bovine serum albumin and detection probes, applied in the field of bovine serum albumin detection probes and its preparation, can solve the problems of harsh detection conditions, cumbersome detection steps, high toxicity of reagents, etc., and achieve low detection cost and excellent preparation method Simple, Widely Applicable Effects

Inactive Publication Date: 2018-06-22
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current detection methods all require expensive instruments, and the detection steps are cumbersome, time-consuming, and the detection conditions are harsh, and the reagents used in the detection are highly toxic

Method used

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  • A bovine serum albumin detection probe and its preparation method and application
  • A bovine serum albumin detection probe and its preparation method and application
  • A bovine serum albumin detection probe and its preparation method and application

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preparation example Construction

[0028] This embodiment provides a method for preparing a bovine serum albumin detection probe, such as figure 1 shown, which includes:

[0029] Step Ⅰ: Mix absolute ethanol and ammonia water at a volume ratio of 2-4:1, then slowly add tetraethyl orthosilicate ethanol solution, stir at 20-30°C for 1 hour, and then mix with 3-aminopropyl triethoxy base silicon mixed, stirred at 20-30°C for 16-24h, dried at 40-55°C to obtain NH 2 -SiO 2 .

[0030] Tetraethyl orthosilicate is hydrolyzed to SiO under the action of ammonia water 2 Nanoparticles. Then react with 3-aminopropyl triethoxy silicon (APTES), in SiO 2 The surface-bonded amino groups give NH 2 -SiO 2 . The purpose of slowly adding TEOS ethanol solution is to control the reaction rate, so that the generated SiO 2 Nanoparticles have a small particle size. By the synthesis method provided by the invention, the obtained SiO 2 The particle size of the bovine serum albumin detection probe is further reduced, which is be...

Embodiment 1

[0051] The present embodiment provides a kind of preparation method of bovine serum albumin detection probe, such as figure 1 shown, which includes:

[0052] First, tetraethyl orthosilicate and absolute ethanol are mixed at a volume ratio of 1:2 to form an ethanol solution of tetraethyl orthosilicate; then anhydrous ethanol and ammonia water are mixed at a volume ratio of 2:1; then slowly add the above prepared Tetraethyl orthosilicate ethanol solution, stirred at 20-30°C for 1h. Then mix with 3-aminopropyl triethoxy silicon, wherein the volume ratio of 3-aminopropyl triethoxy silicon to tetraethyl orthosilicate is 1:12; the above solution is stirred at 20-30 °C 16h, dried at 55°C to give NH 2 -SiO 2 .

[0053] Mix BSA with EDC and NHS, wherein the mass ratio of the three is BSA:EDC:NHS 8:1:1; the above mixed solution is stirred at 20-30°C for 3h. Subsequent with NH 2 -SiO 2 mixed, where NH 2 -SiO 2 The mass ratio to BSA is 1:4; the above solution is stirred at 20-30°...

Embodiment 2

[0055] The present embodiment provides a kind of preparation method of bovine serum albumin detection probe, it comprises:

[0056] First, tetraethyl orthosilicate and absolute ethanol are mixed at a volume ratio of 1:3 to form an ethanol solution of tetraethyl orthosilicate; then anhydrous ethanol and ammonia water are mixed at a volume ratio of 3:1; then slowly add the above prepared Tetraethyl orthosilicate ethanol solution, stirred at 20-30°C for 1h. Then mix with 3-aminopropyltriethoxysilicon, wherein the volume ratio of 3-aminopropyltriethoxysilicon to tetraethyl orthosilicate is 1:14; the above solution is stirred at 20-30°C 20h, dried at 50°C to give NH 2 -SiO 2 ;

[0057] Mix BSA with EDC and NHS, wherein the mass ratio of the three is BSA:EDC:NHS 10:1:1; the above mixed solution is stirred at 20-30°C for 2h. Subsequent with NH 2 -SiO 2 mixed, where NH 2 -SiO 2 The mass ratio to BSA is 1:5; the above solution is stirred at 20-30°C for 2h to obtain BSA-SiO 2 ....

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Abstract

The invention provides a bovine serum albumin detection probe and its preparation method and application. The preparation method uses cheap tetraethyl orthosilicate as a raw material, and the tetraethyl orthosilicate is hydrolyzed under the action of ammonia water to synthesize nano Grade silica particles; then use 3-aminopropyltriethoxysilane to bond amino groups on the surface of silica particles to obtain NH2-SiO2; then bond BSA to NH2-SiO2 to obtain BSA -SiO2. The preparation method is simple, the reaction conditions are mild, easy to realize, and suitable for large-scale industrial production. The detection method using BSA-SiO2 to detect BSA utilizes the specific binding reaction between antigen and antibody, and at the same time uses HRP to label the antibody to achieve visual detection, with strong specificity and high sensitivity.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a detection probe for bovine serum albumin and its preparation method and application. Background technique [0002] In the process of culturing cells to produce vaccines, bovine serum is the main component of the medium. Since bovine serum albumin is often contained in bovine serum, it is very important to detect residual bovine serum albumin (BSA) in vaccines, which is directly related to the safety of biological products. However, the current detection methods all require expensive instruments, and at the same time, the detection steps are cumbersome, time-consuming, and the detection conditions are harsh, and the reagents used in the detection are highly toxic. Contents of the invention [0003] The first object of the present invention is to provide a bovine serum albumin detection probe, which has a small particle size, stable structure and easy preservation. The bovine s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N2333/765
Inventor 青琳森王乾龙丁立生梁健饶亮明李兴德蒋立银帅克
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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