Method for performing in-situ quantitative detection on interaction of gold nanoparticle surface adsorption protein and cytomembrane associated receptor based on quartz crystal chip

A technology of gold nanoparticles and quartz crystals, which is applied in the innovative development of dissipative quartz crystal microbalance detection methods, can solve problems such as the complex structure of nanomaterials-protein coronas, and achieve the effect of strong practicability and simple operation

Active Publication Date: 2017-09-08
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

As for the complex structure of the nanomaterial-protein corona, there is no report on the technical invention of the in situ quantitative detection of the...

Method used

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  • Method for performing in-situ quantitative detection on interaction of gold nanoparticle surface adsorption protein and cytomembrane associated receptor based on quartz crystal chip
  • Method for performing in-situ quantitative detection on interaction of gold nanoparticle surface adsorption protein and cytomembrane associated receptor based on quartz crystal chip
  • Method for performing in-situ quantitative detection on interaction of gold nanoparticle surface adsorption protein and cytomembrane associated receptor based on quartz crystal chip

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Experimental program
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Embodiment 1

[0053] (1) Quartz crystal chip pretreatment:

[0054] The cleaned quartz crystal chip is placed in a 12-well plate, and a mixture of 2-mercaptopropionic acid containing 0.97% volume fraction and 2-mercaptoethanol containing 0.95% volume fraction is added to the 12-well plate, 1ml per hole, After treatment for 12 hours, wash off the small sulfhydryl molecules not bound to the chip with three times of water, and blow dry with nitrogen for use;

[0055] (2) Activation of carboxyl groups on the surface of quartz crystal chips:

[0056] Put the pretreated quartz crystal chip into the QCM-D instrument, first pass water three times at a flow rate of 100 μl / min, wash for 10 minutes, wait until the signal curve is stable, and then pass it with 75 mM EDC and 30 mM EDC at a flow rate of 30 μl / min NHS solution for 1h, wait until the signal curve is stable;

[0057] (3) Preparation and immobilization of gold nanoparticles on chiral surfaces:

[0058] Take 10ml of nano-gold colloid solut...

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Abstract

The invention relates to a method for performing in-situ quantitative detection on interaction of gold nanoparticle chiral surface adsorption protein and a cytomembrane associated receptor based on a quartz crystal chip. The method comprises the following steps: (1) pretreating the quartz crystal chip by 2-mercaptopropionic acid and 2-mercaptoethanol; (2) putting the treated chip into a QCM-D instrument, introducing EDC/NHS and activating the carboxyl on the surface of the chip; (3) introducing gold nanoparticles with the surface modified by penicillamine micromolecules and combining an amido bond and the chip; (4) introducing ligandin and enabling the ligandin to be adsorbed on the chiral surface of the nanogold; (5) introducing cell derived lipidosome with the surface containing the associated receptor. The interaction of the ligandin and the associated receptor is quantified through the intensity of a frequency signal. The method is simple to operate and high in practicability, and has important significance in research on the structure and the function of protein crown formed on the surfaces of the nanoparticles.

Description

technical field [0001] The invention belongs to the innovative development field of detection methods of dissipative quartz crystal microbalances, and mainly relates to a method for in-situ quantitative detection of interactions between proteins adsorbed on the chiral surface of gold nanoparticles and cell membrane-related receptors based on quartz crystal chips. Background technique [0002] Dissipative quartz crystal microbalance (QCM-D) utilizes the piezoelectric characteristics of the quartz crystal resonator to convert the surface quality change of the quartz crystal chip electrode into the frequency change of the output electrical signal of the quartz crystal oscillator circuit, and then obtain high-precision balance through the computer. Data is a new type of sensitive quality detection instrument, and its measurement accuracy can reach nanogram level. [0003] When the nanomaterial enters the human body, due to its large specific surface area, it will absorb the prot...

Claims

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Application Information

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IPC IPC(8): G01N27/26
CPCG01N27/26
Inventor 陈春英王明哲王新一
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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