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Preparation method of stable and uniform nanopore

A nano-pore and stable technology, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of uneven pore size, high difficulty of reproduction, unstable structure, etc., and achieve the effect of stable pore structure

Active Publication Date: 2022-04-19
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, the large-aperture nanopores formed by natural nanopores, especially the self-assembly of protein monomers, often have unstable structures and uneven pore sizes, which lead to problems such as low success rate of experiments and high difficulty in reproduction, which seriously affect their practical detection. application potential in

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  • Preparation method of stable and uniform nanopore
  • Preparation method of stable and uniform nanopore
  • Preparation method of stable and uniform nanopore

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Embodiment Construction

[0023] The concentration of the experimental reagents and the duration of the experiment involved in the following specific examples can be adjusted according to the application purpose and the change of the natural protein material. The basic idea of ​​the present invention is described for the implementation objects, and the following embodiments and the features in the embodiments can be combined with each other under the condition of no conflict.

[0024] Such as figure 1 As shown, this embodiment relates to a method for manufacturing a large-aperture nanochannel based on an engineered natural pore-forming protein. The protein used in this embodiment is a mutant of the cholesterol-dependent bacterial toxin protein PFO. The specific method includes the following steps :

[0025] Step 1) Purification of PFO mutant monomeric protein:

[0026] For the G57 and S190 of the PFO monomer sequence (that is, the glycine at the 57th position of the protein sequence and the threonine...

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Abstract

The invention relates to a preparation method of stable and uniform nanopores, which comprises the following steps: mixing and incubating a nanopore monomer and a lipid vesicle physiological buffer turbid liquid with uniform particle size, and forming a uniform and stable nanopore precursor on lipid vesicles; sequentially adding an unlocking agent to enable the nanopore precursor on the lipid vesicles to form nanopores, adding a cross-linking agent to enable the formed nanopore structure to be more stable, and adding a surfactant to destroy the lipid vesicles and enable the nanopores to enter the aqueous solution to form a mixed solution of the nanopores and lipid; and separating and purifying to obtain the nanopores with consistent molecular weight, uniform pore diameter and stable structure. The pore-forming protein obtained by the invention can form a pre-pore-forming compound on specially-made lipid vesicles, and then a cross-linking agent and a surfactant are combined, so that the structure of a nanopore is more stable, and the pore-forming success rate is higher.

Description

technical field [0001] The invention relates to a technology in the field of biological nanomaterials, in particular to an engineered method for preparing nanopores with a stable structure and a uniform pore diameter greater than 3 nanometers. Background technique [0002] The nanopore sensor separates the two chambers containing the electrolyte through a lipid bimolecular membrane, etc., and the nanopore is embedded in the separation material to connect the two chambers, and the ions will pass freely under the action of an electric bias. The pores generate a constant opening current; and when the molecular substances in the biological solution enter the nanopore due to the electrophoretic force, and the nanopore is partially blocked through the hole, the flow of ions in it will be partially hindered, and the degree of obstruction is the same as that of the biomolecules entering the nanopore. The size and shape of the substance in the pore are closely related, and are also a...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/26
CPCG01N27/3275G01N27/3278G01N27/26
Inventor 阮鉴鉴刘美君于晗丹尼尔·恰可夫邵志峰程酩沈轶
Owner SHANGHAI JIAO TONG UNIV
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