Method for quantitatively detecting ampicillin by marking antibody with laterally assembled gold nanorod

A technology of ampicillin and gold nanorods is applied in the field of immunodetection to achieve the effects of being conducive to sensitive detection, simple sample pretreatment and good stability

Inactive Publication Date: 2012-10-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far in China, there is no report on the detection of in vitro targets using the signal of the laser particle size analyzer of gold nanorods. The present invention makes up for this gap.

Method used

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  • Method for quantitatively detecting ampicillin by marking antibody with laterally assembled gold nanorod
  • Method for quantitatively detecting ampicillin by marking antibody with laterally assembled gold nanorod
  • Method for quantitatively detecting ampicillin by marking antibody with laterally assembled gold nanorod

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 The coupling of ampicillin antibody and coating agent to gold nanorods:

[0031] a. Coupling of antibody and gold nanorods: 0.5mL, 100μg / mL ampicillin antibody was added dropwise to 0.5mL, 2nmol / L gold nanorod solution pre-adjusted to pH 8.5-8.8 with 0.1mol / L potassium carbonate , shake and mix at room temperature, react at 37°C for 1 h, centrifuge at 6000 rpm for 20 min after the reaction, remove unbound antibody, resuspend in pH 7.4, 0.01 mol / L PBS (containing 0.5% PEG20000), and prepare ampicillin antibody-modified Gold nanorod probe;

[0032] b. Coupling agent and gold nanorod coupling: the method is similar to that of antibody and gold nanorod coupling, 0.5mL, 100μg / mL ampicillin coating agent is added dropwise and pre-adjusted to pH 5 with 0.1mol / L hydrochloric acid , 0.5mL, 2nmol / L gold nanorod solution, shake and mix at room temperature, react at 37°C for 1h, centrifuge at 6000rpm for 20min at the end of the reaction, remove unbound coating origina...

Embodiment 2

[0033] Embodiment 2 laser particle size analyzer detection

[0034] Add 10 μL modified ampicillin-coated original gold nanorod solution and 10 μL ampicillin standard of different concentrations into a 1.5 mL centrifuge tube, mix well, then add 10 μL modified ampicillin antibody gold nanorod solution, vortex to mix After incubating at 37°C for 0.5h, take 20μL of the reaction solution and dilute it to a volume of 1mL, add it to the plastic detection cuvette, and measure the size of the immune polymer assembled on the side of the gold nanorod after the reaction. Ampicillin concentration C standard curve Size ~ C.

[0035]The solutions used were PBS buffer solution with pH 7.4 and 0.01 mol / L; the concentrations of ampicillin standard were 0, 0.1, 0.5, 1.0, 5.0, 10.0, 20.0, 100 ng / mL.

Embodiment 3

[0036] Embodiment 3 Quantitative detection by laser particle size analyzer

[0037] Measure the particle size of the gold nanorods with the Zetasizer Nano ZS system of the British Malvern laser particle size analyzer,

[0038] The detection parameters used in this experiment are: the detection temperature is 20°C, the detection angle is 173°, the excitation light source wavelength is 633nm, and the laser power is 5mW.

[0039] The principle of side assembly detection of gold nanorods: ampicillin antibody and coating original are modified on the side of gold nanorods, and ampicillin standard or supernatant to be tested is added, and ampicillin standard and coating original are competitively modified on gold nanorods. The antibody on the rod, with the amount of ampicillin standard added, the size of the formed antibody is different from that of the immune polymer assembled on the side of the original gold nanorod, which is detected by a laser particle size analyzer Detection of...

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Abstract

A method for quantitatively detecting ampicillin by marking an antibody with a laterally assembled gold nanorod belongs to the technical field of immunodetection. In the invention, an ampicillin antibody and primordial covering are decorated on the lateral side of a gold nanorod, then ampicillin standard substance or supernate to be detected are added, the ampicillin standard substance competes with the primordial covering for the antibody decorated on the lateral side of the gold nanorod, as the additions of ampicillin standard substance differentiate, the formed grain diameters of immune polymers assembled on the lateral side of the gold nanorod by the antibody and the primordial covering have different sizes, thus the detection of ampicillin-containing sample to be detected can be finished by using a laser granularity instrument. The invention dispenses with complicated sample pre-treatment, has convenient operation, namely, after hatching, the sample to be detected can be directlydetected with the instrument in one step; the gold nanorod for marking is 13*40nm, has high water-solubility and good dispersity and stability.

Description

technical field [0001] The invention relates to a method for quantitatively detecting ampicillin with a side-assembled gold nanorod-labeled antibody, which belongs to the technical field of immunodetection. Background technique [0002] Ampicillin is a semi-synthetic penicillin, which introduces an amino group at the alpha position of the carboxyl group of the side chain of penicillin G, which changes its polarity, making it easier to penetrate the bacterial cell membrane and expand the antibacterial spectrum. The structure is: [0003] [0004] Ampicillin is acid-resistant and not enzyme-resistant. It is easily absorbed by oral administration or intramuscular injection. It is not as effective as penicillin against most Gram-positive bacteria. The characteristic is that it has a strong effect on Gram-negative bacteria, such as Escherichia coli, Proteus, and Salmonella. At the same time, it has the characteristics of acid resistance, which avoids the disadvantage that nat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/531G01N15/02
Inventor 胥传来朱颖越陈伟徐丽广马伟彭池方
Owner JIANGNAN UNIV
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