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CN-119688896-B - Cell membrane bionic nano-recognition material and preparation method and application thereof

CN119688896BCN 119688896 BCN119688896 BCN 119688896BCN-119688896-B

Abstract

The invention provides a cell membrane bionic nano recognition material, a preparation method and application thereof, wherein magnetic nano particles are adopted as carriers, cell membranes are fixed on the magnetic carriers through biological orthogonal reaction, the cell membrane-based bionic nano recognition material is finally prepared and is used for recognizing unknown pollutants in foods and evaluating risks, the characteristics of high magnetic response of the magnetic nano particles and high complex biological interface performance mediated by the cell membranes are combined, and active cell membranes containing target receptors are wrapped on the surfaces of the magnetic nano particles in a covalent fixation manner, so that the integrity of the cell membranes and the three-dimensional structure and biological activity of membrane receptor proteins on the cell membranes are maintained to the greatest extent, excellent targeting performance of the bionic nano material is provided, the specific screening and rapid magnetic separation of unknown chemical risks in complex food matrixes are realized, the recognition efficiency of the unknown chemical risks in foods is improved, and a novel concept with toxicity as a guide is provided for screening of potential pollutants in foods.

Inventors

  • LI GUOLIANG
  • LI QIANYU
  • LIU JIANGHUA
  • LI WENRUI

Assignees

  • 陕西科技大学

Dates

Publication Date
20260512
Application Date
20241219

Claims (6)

  1. 1. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples is characterized by comprising the following steps of preprocessing a sample to be detected to obtain a food sample extracting solution, adding the cell membrane bionic nano-recognition material into the food sample extracting solution, carrying out vortex mixing, collecting the material through a magnet, eluting by adopting a mixed solution of isopropanol and water, and finally analyzing an eluent by using a high performance liquid chromatography tandem time-of-flight mass spectrum to screen out beta-carboline and norbeta-carboline, wherein the preprocessing comprises the steps of cutting the sample to be detected into blocks, carrying out homogenization treatment, adding deionized water and acetonitrile into the homogenized sample, carrying out vortex treatment and ultrasonic treatment, then adding anhydrous magnesium sulfate, sodium chloride and n-hexane, continuing vortex mixing, centrifuging again after the sample fully acts with the solvent, collecting an intermediate acetonitrile layer, adding n-hexane again to remove residual lipid, and finally, carrying out reduced pressure distillation and drying the degreasing solution, and redissolving by using deionized water and methanol to complete the preprocessing; the cell membrane bionic nano-recognition material is prepared by the following method: S1, preparing Fe 3 O 4 magnetic nano particles, coating SiO 2 on the magnetic nano particles to obtain Fe 3 O 4 @SiO 2 , and then adding 3-aminopropyl triethoxysilane for amination modification to obtain Fe 3 O 4 @SiO 2 -NH 2 ; S2, modifying the Fe 3 O 4 @SiO 2 -NH 2 by using DBCO-NHS to obtain dibenzocyclooctyne functionalized magnetic nano particles, culturing HEK293 cells which highly express human epidermal growth factor receptor, marking azide groups on the cell surface by non-natural sugar metabolism sugar engineering during the culture period, collecting cells in a logarithmic phase, and obtaining azide modified cell membranes by ultrasonic centrifugation; S3, performing biological orthogonal reaction on the cell membrane subjected to azide modification and the dibenzocyclooctyne functionalized magnetic nano-particles to prepare the cell membrane bionic nano-recognition material; The Fe 3 O 4 magnetic nano-particles are prepared by adding ferric trichloride hexahydrate, ammonium acetate and sodium citrate into ethylene glycol, stirring at room temperature for reaction, and then performing heat treatment on the reaction product to obtain the Fe 3 O 4 magnetic nano-particles, wherein the mass ratio of the ferric trichloride hexahydrate to the sodium citrate is (3-6) (1-2), the heat treatment temperature is 180-220 ℃ and the time is 16-17 h; The Fe 3 O 4 @SiO 2 -NH 2 is prepared by adding Fe 3 O 4 @SiO 2 into toluene, adding 3-aminopropyl triethoxysilane after ultrasonic dispersion, and stirring for reaction to obtain Fe 3 O 4 @SiO 2 -NH 2 , wherein the ratio of Fe 3 O 4 @SiO 2 to 3-aminopropyl triethoxysilane is (0.1-0.2) g (1-2) mL.
  2. 2. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples, which is disclosed in claim 1, is characterized in that the preparation of Fe 3 O 4 @SiO 2 comprises the steps of adding Fe 3 O 4 magnetic nano-particles into ethyl silicate, adding ammonia water, ethanol and water, and stirring for reaction to obtain Fe 3 O 4 @SiO 2 , wherein the ratio of the Fe 3 O 4 magnetic nano-particles to the ethyl silicate is (0.1-0.2) g (0.8-1.6) mL.
  3. 3. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples, which is characterized in that the preparation of the dibenzocyclooctane functionalized magnetic nano-particles is characterized in that Fe 3 O 4 @SiO 2 -NH 2 is added into DBCO-NHS solution, room temperature reaction is carried out, the dibenzocyclooctane functionalized magnetic nano-particles are prepared, the ratio of Fe 3 O 4 @SiO 2 -NH 2 to DBCO-NHS is (0.1-0.2) g (3-6) mL, and the concentration of DBCO-NHS is 4.45 mM.
  4. 4. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples according to claim 1, wherein the cell surface is marked with azide groups through non-natural sugar metabolism sugar engineering, specifically, ac 4 ManNAz is added into a complete culture medium for fully mixing to culture EGFR-HEK293 cells, when the cells grow to a logarithmic phase, trypsin digestion liquid is adopted for digestion and collection, centrifugation and ultrasonic disruption are carried out, and phosphate buffer solution is added for resuspension, so that the cell membrane with the azide modification is prepared.
  5. 5. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples, which is characterized in that the cell membrane bionic nano-recognition material is prepared by performing biological orthogonal reaction on the azide-modified cell membrane and the dibenzocyclooctane functionalized magnetic nano-particles, specifically, the cell membrane bionic nano-recognition material is prepared by adding the dibenzocyclooctane functionalized magnetic nano-particles into a suspension of an azide-modified cell membrane after ultrasonic dispersion, and performing vibration mixing reaction at room temperature, wherein the mass ratio of the dibenzocyclooctane functionalized magnetic nano-particles to the azide-modified cell membrane is 1 (2.5-3).
  6. 6. The application of the cell membrane bionic nano-recognition material in screening potential chemical risks in food samples according to claim 1, wherein the ratio of the cell membrane bionic nano-recognition material to the food sample extracting solution is (5-10) mg (1-2) mL.

Description

Cell membrane bionic nano-recognition material and preparation method and application thereof Technical Field The invention belongs to the technical field of material detection, and relates to a cell membrane bionic nano-recognition material, a preparation method and application thereof. Background With the continued sophistication of food regulatory systems, food safety concerns have presented a trend toward the shift from known contaminant detection to unknown risk identification. Overtreatment of the food product, package contamination and additive use may all lead to unpredictable risk generation. The compounds often have complex structures, have chemical risk hidden dangers which are not recognized yet, lack relevant safety standard regulations and form a great threat to public health. Thus, accurate identification of potentially risky compounds in food is critical to food safety monitoring and risk assessment. Currently, high-performance liquid chromatography tandem mass spectrometry (HPLC-MS) is widely applied to screening of illegal addition of chemical pollutants such as agricultural and veterinary drugs and metabolites thereof. However, identifying unknown compounds remains a challenging task, particularly without relevant guidance. Thus, identification of potential contaminants in complex food samples requires poison-directed screening strategies. In recent years, toxicity evaluation of contaminants has been mainly based on cell or biological experiments, and belongs to a conventional non-targeted screening method, in which samples are comprehensively analyzed to find various chemical substances possibly existing therein without presetting specific target compounds. Since the non-targeted screening does not presets the target compound, it is suitable for comprehensive analysis of unknown or complex samples. However, because of the complexity of intracellular processes, non-targeted screening makes it difficult to accurately identify specific compounds of interest and their mechanisms of action, which makes elucidation of specific targets and their mechanisms of action difficult. In addition, non-targeted screening typically requires a large number of sample and experimental resources, is costly, takes a long time, and results may be affected by a variety of factors, such as experimental conditions, matrix interference, etc., making it less accurate. Disclosure of Invention Aiming at the problems in the prior art, the invention provides a cell membrane bionic nano identification material and a preparation method and application thereof, thereby solving the technical problems that a specific target compound and an action mechanism thereof are difficult to accurately identify, the cost is high, the time consumption is long and the accuracy is low when non-targeted screening of potential risk substances in complex food matrixes is utilized in the prior art. The potential risk in the complex food matrix is found to be a technical problem of high cost and long time consumption. The invention is realized by the following technical scheme: A preparation method of a cell membrane bionic nano-recognition material comprises the following steps: S1, preparing Fe 3O4 magnetic nano particles, coating SiO 2 on the magnetic nano particles to obtain Fe 3O4@SiO2, and then adding 3-aminopropyl triethoxysilane for amination modification to obtain Fe 3O4@SiO2-NH2; S2, modifying the Fe 3O4@SiO2-NH2 by using DBCO-NHS to obtain dibenzocyclooctyne functionalized magnetic nano particles, culturing HEK293 cells which highly express human epidermal growth factor receptor, marking azide groups on the cell surface by non-natural sugar metabolism sugar engineering during the culture period, collecting cells in a logarithmic phase, and obtaining azide modified cell membranes by ultrasonic centrifugation; s3, performing biological orthogonal reaction on the cell membrane subjected to azide modification and the dibenzocyclooctyne functionalized magnetic nano-particles to obtain the cell membrane bionic nano-recognition material. Preferably, the Fe 3O4 magnetic nano-particles are prepared by adding ferric trichloride hexahydrate, ammonium acetate and sodium citrate into ethylene glycol, stirring at room temperature for reaction, and then performing heat treatment on a reaction product to obtain the Fe 3O4 magnetic nano-particles, wherein the mass ratio of the ferric trichloride hexahydrate to the sodium citrate is (3-6) (1-2), the heat treatment temperature is 180-220 ℃ and the time is 16-17 h. Preferably, the Fe 3O4@SiO2 is prepared by adding Fe 3O4 magnetic nano particles into ethyl silicate, adding ammonia water, ethanol and water, and stirring for reaction to obtain Fe 3O4@SiO2, wherein the ratio of the Fe 3O4 magnetic nano particles to the ethyl silicate is (0.1-0.2) g (0.8-1.6) mL. Preferably, the Fe 3O4@SiO2-NH2 is prepared by adding the Fe 3O4@SiO2 into toluene, adding 3-aminopropyl triethoxysilane after ultrasonic dispers