Microbial nano-enzyme probe, test strip for detecting salbutamol and application of test strip

A salbutamol and nano-enzyme technology, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of waste, difficulty in obtaining probes, etc. simple effect

Pending Publication Date: 2022-05-20
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, during the preparation of the probe, due to the IrO 2 The extremely small particle size of NPs makes it difficult to obtain intact probes by high-speed centrifugation, resulting in a large amount of monoclonal antibodies (mAbs) being wasted

Method used

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  • Microbial nano-enzyme probe, test strip for detecting salbutamol and application of test strip
  • Microbial nano-enzyme probe, test strip for detecting salbutamol and application of test strip
  • Microbial nano-enzyme probe, test strip for detecting salbutamol and application of test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1: the preparation of the probe of the test strip of rapid detection albuterol

[0062] combine figure 1 According to the above technical scheme, this example provides microbial nanozyme probes and preparation methods, including yeast cells loaded with iridium oxide nanoparticles as signal carriers, and then adding salbutamol monoclonal antibodies for adsorption. Include the following steps:

[0063] (1) Preparation of iridium oxide nanoparticles (IrO 2 NPs): mix 50mg trisodium citrate and 30mg K 2 IrCl 6 Mix with 50 mL of ultrapure water, adjust the pH of the solution to 7.5, and reflux under boiling. When the color of the solution changed from brown to steel gray, the solution was continued to be stirred and heated for 30 minutes. After the solution was cooled to room temperature, the pH was readjusted to 7.5, and then the solution was heated for 30 minutes until the pH was stable, and the color of the final solution turned dark blue to obtain IrO 2 ...

Embodiment 2

[0068] Embodiment 2: Preparation of the immunochromatographic test strip for rapid detection of albuterol

[0069] This embodiment provides a high-sensitivity immunochromatographic test strip for rapid detection of albuterol, including a nitrocellulose membrane, a sample pad, a conjugation pad, an absorbent pad and a backing plate, on which a nitrocellulose membrane, nitrocellulose One end of the membrane is covered with a water-absorbing pad, and the other end of the nitrocellulose membrane is covered with a sample pad and a binding pad in turn. The non-covered surface of the nitrocellulose membrane is provided with a detection line and a control line along the horizontal direction, and the binding pad and the sample pad are respectively sealed with a blocking solution. deal with.

[0070] The preparation method of the nitrocellulose membrane comprises: 1 mg / mL salbutamol-bovine serum albumin conjugate is coated on the detection line at the streaking rate of 0.8 μ L / cm to obt...

Embodiment 3

[0075] Embodiment 3: the optimization of the test strip that detects albuterol rapidly

[0076] For best detection performance, the system optimizes the Yeast@IrO 2 -Some relevant experimental parameters of ICA. In a typical competitive immunoassay, the amount of mAbs (2, 3, 4, 5 and 6 μg), the amount of T-line streak (1, 0.9, 0.8, 0.7, 0.6 and 0.5 μL / cm), the amount of probe (3, 4, 5, 6 and 7 μL) and immunization time (5, 10, 15 and 20 min) directly affected the sensitivity and visibility of the proposed biosensor. According to the signal intensity of the T line and the competitive inhibition rate, the optimal parameters of this biosensor were selected as the main criteria. The calculation formula of competitive inhibition rate is (1-T / T 0 )×100%, where T 0 and T are the T-line intensities of SAL-negative and SAL-positive solutions, respectively.

[0077] See image 3 A, As the number of anti-SAL-mAbs increased, the intensity of T lines increased significantly. Meanwhi...

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Abstract

The invention discloses a microbial nano-enzyme probe, a method for detecting salbutamol and application, the microbial nano-enzyme probe comprises a salbutamol monoclonal antibody and a signal carrier, and the signal carrier is yeast cells loaded with iridium oxide nanoparticles. According to the present invention, the iridium oxide nanoparticle-loaded yeast cells are adopted to construct the probe during the immunochromatography test strip detection for the first time, the [3, 3 ', 5, 5'-tetramethyl benzidine (TMB) and the hydrogen peroxide (H2O2)] are adopted as the substrates to trigger the chromogenic reaction, the high-sensitivity target object detection can be achieved, the detection limit is as low as 0.012 ng / mL, and the sensitivity is improved by 55 times compared with the sensitivity of the traditional gold-labeled test strip; in addition, the method has a good application prospect, and a universal platform can be provided for sensitively, accurately and rapidly detecting the food-borne stimulant. Meanwhile, the salbutamol detection kit can be successfully applied to salbutamol detection in pork liver and beef samples.

Description

technical field [0001] The invention belongs to the field of biological detection, and relates to a microbial nanozyme probe, a method for detecting salbutamol and its application, and specifically relates to a yeast cell loaded with iridium oxide nanoparticles as a signal carrier, and the probe is prepared by labeling an antibody, and the probe is used The test strip, and its application for rapid and sensitive detection of albuterol. Background technique [0002] Food-borne stimulants in food are mainly β 2 - Receptor agonists, eating foods containing these food-derived stimulants will cause harm to people's body and psychology, and is very detrimental to the healthy development of people's mind and body. Salbutamol (SAL) is a fast-acting beta 2 - Adrenoceptor agonists that selectively stimulate beta of bronchial smooth muscle 2 In terms of clinical treatment, it is used to treat bronchial asthma. However, SAL absorbs quickly in the body and is easy to accumulate in th...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/577G01N33/558G01N33/543G01N33/53G01N33/52
CPCG01N33/587G01N33/577G01N33/558G01N33/54346G01N33/5308G01N33/52Y02A50/30
Inventor 王丽赵爽何坤益补彤白菲儿杨铠溶
Owner NORTHWEST A & F UNIV
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