WO2022094912A1 - Sonde biologique, son procédé de fabrication et son application - Google Patents

Sonde biologique, son procédé de fabrication et son application Download PDF

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WO2022094912A1
WO2022094912A1 PCT/CN2020/127031 CN2020127031W WO2022094912A1 WO 2022094912 A1 WO2022094912 A1 WO 2022094912A1 CN 2020127031 W CN2020127031 W CN 2020127031W WO 2022094912 A1 WO2022094912 A1 WO 2022094912A1
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solution
group
gold
sialic acid
substituted
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PCT/CN2020/127031
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Chinese (zh)
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李伟
王怀雨
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深圳先进技术研究院
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses

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  • the invention belongs to the technical field of biochemistry, and relates to a biological probe and a preparation method and application thereof.
  • the pathogen causing new coronary pneumonia is a new type of coronavirus (referred to as "new coronavirus").
  • the detection methods of 2019-nCoV mainly include nucleic acid RNA detection and 2019-nCoV IgG/IgM antibody detection.
  • the former extracts nucleic acid substances from the sample to be tested and uses polymerase chain reaction (PCR) to amplify the RNA of the sample to be tested.
  • PCR polymerase chain reaction
  • the real-time quantitative PCR technology is generally used for detection; the latter uses that after a person is infected with the new coronavirus, the immune system will react to produce corresponding IgG and IgM antibodies, by detecting the new coronavirus in human blood.
  • Viral IgG and IgM are used to indirectly indicate whether the subject has a new coronavirus infection, and the colloidal gold method is generally used to detect these two antibodies.
  • Nucleic acid RNA detection generally uses RT-PCR technology to directly detect whether there is new coronavirus RNA in the sample, but it needs to manually extract the RNA virus in the sample, which is affected by multiple factors such as sample sampling method, sample transportation and storage, manual operation, etc., often accompanied by false negative interference , and a small number of people with positive RNA test results did not show infectivity, so there was also false positive interference;
  • the new coronavirus IgG/IgM antibody test is an indirect detection method, by detecting whether there is a new coronavirus IgG/IgM antibody in the blood. To indirectly characterize whether the subject is infected with the new coronavirus, but it cannot indicate whether there are new coronavirus particles in the subject in real time, so it is impossible to judge whether the subject is infectious.
  • the purpose of the present invention is to provide a biological probe and its preparation method and application.
  • the present invention provides a biological probe comprising a probe carrier and a customized sialic acid ligand molecule connected to the probe carrier;
  • R 1 is selected from hydroxyl, methoxy or substituted methoxy
  • R 2 is selected from acetamido, substituted acetamido, benzamido, substituted benzoyl, alkoxycarbonamido, triazole or substituted triazolyl
  • R is selected from hydroxyl, methoxy, substituted methoxy, acetamido, substituted acetamido, sulfonamide or phosphoramido
  • A is n is selected from 0, 1, 2, 3, 4, 5, 6, 7, and m is selected from 2, 3, 4, 5, 6, 7, 8, 9.
  • the customized sialic acid ligand molecule can be further matched to occupy the sugar-binding domain pocket of the new coronavirus S protein through the derivatization of R 1 , R 2 and R 3 groups, and has high specificity and binding force to the new coronavirus S protein.
  • substituted methoxy group is X 1 -CH 2 O-, wherein X 1 is selected from phenyl or vinyl;
  • the substituted acetamido is X 2 -CH 2 CONH-, wherein X 2 is selected from methyl, ethyl, n-propyl, isopropyl, hydroxymethyl or hydroxyethyl;
  • the substituted benzamido is X 3 -BzNH-, wherein the number of X 3 is at least 1 and is located at any position of the benzene ring, and X 3 is selected from at least one of halogen atoms, methyl groups, methoxy groups, and nitro groups.
  • the alkoxycarbonamide group is X 4 -OC(O)NH-, wherein X 4 is selected from benzyl, allyl, tert-butyl or trichloroethyl;
  • the substituted triazolyl is wherein X 5 is selected from substituted phenylalkyl (X 3 -Ph-(CH 2 ) n1 -) or substituted carbonyl (X 6 -C(O)-), wherein X 3 is at least 1 in number and located in the benzene Any position of the ring, X 3 is selected from at least one of halogen atom, methyl group, methoxy group and nitro group, n1 is selected from 0, 1, 2; X 6 is selected from alkoxy group CH 3 -(CH 2 ) n2 -O- or alkylamino CH3- ( CH2 ) n2 -NH-, n2 is selected from 0,1,2,3,4,5.
  • the probe carrier is selected from nanomaterials with sensitive LSPR effect or substances with obvious signal changes in a dispersed-aggregated state;
  • the nanomaterials with sensitive LSPR effect include gold-based nanoparticles and composite nanoparticles with a metallic gold layer on the surface; more preferably, the gold-based nanoparticles include gold nanorods and gold nanospheres;
  • the substance with obvious signal change in the dispersion-aggregation state comprises an aggregation-induced luminescence AIE material;
  • the aggregation-induced luminescence AIE material comprises a tetrathienylthiophene compound with a cyclic polyene skeleton, a cyano-substituted dithiophene Compounds of styrene skeleton, tetrastyrene-type compounds, divinylanthracene-type compounds, tristyryl-type compounds, etc.
  • the molecular end of the aggregation-induced luminescence AIE material has a maleimide-derived group that is reactively linked with a sulfhydryl group.
  • the probe carrier is a gold nanorod
  • the surface of the gold nanorods is coated with a bilayer of cetyltrimethylammonium bromide
  • the gold nanorods have a diameter of 20-25 nm, a length of 60-80 nm, and an aspect ratio of 2.5:1 to 3.5:1.
  • the molar ratio of the customized sialic acid ligand molecules to the gold nanorods is 1 ⁇ 10 4 :1 ⁇ 2 ⁇ 10 4 :1.
  • the present invention provides a preparation method of any of the above-mentioned biological probes, comprising the following steps:
  • the probe carrier solution is a solution of gold nanorods, and the surface of the gold nanorods is coated with a bilayer of cetyltrimethylammonium bromide; the diameter of the gold nanorods is 20-25 nm, and the length is 60-80nm; customized sialic acid ligand molecules are immobilized on the surface of gold nanorods by forming S-Au bonds on the surface of gold nanorods;
  • the molar ratio of the customized sialic acid ligand molecules to the gold nanorods is 1 ⁇ 10 4 :1 ⁇ 2 ⁇ 10 4 :1.
  • the preparation method of the gold nanorod solution is as follows: preparing gold nanorods by a gold seed growth method in an aqueous phase, first reducing chloroauric acid with sodium borohydride to prepare a 1-2 nm gold nanoseed solution; The gold nanoseed solution was added to the mixture of chloroauric acid HAuC1 4 solution, silver nitrate AgNO 3 solution, dilute hydrochloric acid HCl, ascorbic acid solution, and cetyltrimethylammonium bromide CTAB solution. ⁇ 25nm, 60 ⁇ 80nm long, gold nanorod solution with surface covered by cetyltrimethylammonium bromide;
  • the molar ratio of the sodium borohydride and the chloroauric acid is 2:1 to 3:1, the time for the sodium borohydride to reduce the chloroauric acid is 5 to 20 minutes, and the sodium borohydride to reduce the chloroauric acid.
  • the temperature is 25 ⁇ 35 °C;
  • the concentration of the chloroauric acid HAuC1 4 solution is 4.5-5.5 mM, preferably 5 mM, the concentration of the silver nitrate AgNO 3 solution is 0.09-0.12 M, preferably 0.1 M, and the concentration of the dilute hydrochloric acid HCl is 1.0- 1.5mM, preferably 1.2mM, the concentration of the ascorbic acid solution is 8-15mM, preferably 10mM, the concentration of the cetyltrimethylammonium bromide CTAB solution is 0.15-0.25M; the gold nanoseeds
  • the volume ratio of solution, chloroauric acid solution, silver nitrate solution, dilute hydrochloric acid, ascorbic acid solution and cetyltrimethylammonium bromide solution is 45 ⁇ 55 ⁇ l: 5.5 ⁇ 6.5mL: 70 ⁇ 80 ⁇ L: 70 ⁇ 80 ⁇ L: 3.6 ⁇ 3.8mL: 25 ⁇ 35mL, the temperature of the standing reaction is 25 ⁇ 40°C, and the time of the standing reaction is 6 ⁇ 24
  • sialic acid As the starting material, through the protection of the carboxyl group, the R4 protecting group is introduced into the carboxyl group to obtain Generation of peracetylated 2-chlorosugars in the next step of chlorination and acetylation reagents Then the functional side chain precursor is derived from the glycosylation reaction at the 2-position of the sugar ring After deacetylation and carboxyl protection, an intermediate with all the exposed amino and hydroxyl groups on the sugar ring is obtained According to the sequence of R 2 ⁇ R 1 ⁇ R 3 , the introduction of groups is carried out to obtain Finally, through the conversion of side chain groups and the removal of sugar ring protecting groups, the customized sialic acid sugar ligand I was synthesized;
  • the R4 protecting group is introduced into the carboxyl group to obtain Generation of peracetylated 2-chlorosugars in the next step of chlorination and acetylation reagents
  • the functional side chain precursor is derived from the glycosylation reaction at the 2-position of the sugar ring
  • an intermediate with all the exposed amino and hydroxyl groups on the sugar ring is obtained
  • the introduction of groups is carried out to obtain
  • the custom sialic acid sugar ligand I was synthesized through the conversion of side chain groups and the removal of sugar ring protecting groups;
  • R is selected from methyl or benzyl ;
  • a 1 is selected from or -(CH 2 ) m -R 5 ;
  • R 5 is selected from benzyloxy, 2-naphthylmethoxy or allyloxy, n is 0, 1, 2, 3, 4, 5, 6, 7, m are 2,3,4,5,6,7,8,9;
  • PG is the abbreviation of protection group protection group
  • R1 when R1 is hydroxyl, it is sialic acid
  • the group on the molecule does not need to introduce the relevant reaction process of the R1 group;
  • R2 is acetamido, it is sialic acid
  • the group on the molecule does not need to introduce the relevant reaction process of the R2 group;
  • R3 is hydroxyl, it is sialic acid
  • the group on the molecule does not need to introduce the relevant reaction process of the R3 group.
  • the present invention provides an application of any of the above-mentioned biological probes in a novel coronavirus detection reagent.
  • the method of using biological probes to detect the new coronavirus includes the following steps: mixing the biological probes with the sample to be tested, mixing evenly, and then standing for 15-30 minutes, and detecting the new coronavirus by observing the color of the mixed system;
  • the method for using biological probes to detect the new coronavirus specifically includes the following steps: 1) Take two identical aqueous solutions of biological probes and label them as solution C (Control solution, C solution) and solution T (Test solution, T solution), add the control solution to the solution C, add the sample to be tested to the solution T, mix well and let stand for 15-30 minutes to observe the results; 2) Result judgment: positive result: solution C does not change color, solution T changes color; negative Result: solution C does not change color, solution T does not change color; invalid result: solution C changes color, no matter whether solution T changes color or not, another biological probe aqueous solution should be taken for re-testing;
  • the biological probe is a custom sialic acid ligand molecule-gold nanorod biological probe
  • the concentration of the customized sialic acid ligand molecule-gold nanorod biological probe aqueous solution is 10-100 ⁇ M;
  • the volume ratio of the sample to be tested and the customized sialic acid ligand molecule-gold nanorod biological probe aqueous solution is 1:5 to 1:20;
  • the detection limit of the new coronavirus in the sample to be tested is 104 /mL;
  • the sample to be tested is saliva, nasopharyngeal swab, blood, and skin secretions;
  • control solution is deionized water.
  • the present invention provides a biological probe, which can be used to directly detect new coronavirus particles, and has the following beneficial effects compared with the existing new coronavirus and related virus detection methods:
  • the biological probe provided by the present invention directly detects virus particles based on the recognition and binding of sialic acid ligands and S protein on the surface of new coronavirus particles;
  • the biological probe provided by the present invention is used for the detection of the new coronavirus through parallel reactions, and it only takes about 15 minutes to visually judge by color reaction;
  • the biological probe provided by the present invention does not involve multiple procedures such as sample RNA extraction and PCR. It can be directly sampled and then added dropwise, which simplifies the detection structure and avoids manual operation errors. interference caused;
  • the biological probe provided by the present invention is a method for directly detecting new coronavirus particles, which can immediately characterize whether there are new coronavirus particles in the sample;
  • the present invention performs detection by recognizing and combining the sialic acid small molecule with the surface S protein of the new coronavirus, and the sialic acid detection small molecule has obvious advantages such as good economy, high stability, and mass preparation;
  • the present invention provides a biological probe, which improves the recognition and binding of the target protein by modifying the structure of the sialic acid ligand molecule on the surface of the probe. Compared with the natural sugar ligand detection molecule, the detection efficiency is greatly improved. specificity and sensitivity.
  • Fig. 1 is a schematic diagram of the method for detecting new coronavirus particles of the present invention, and the change diagram of the optical signal of the probe LSPR after it becomes a black and white picture is not obvious, wherein the left picture is tea red, and the right picture is dark gray;
  • FIG. 2 is a schematic diagram of a biological probe provided by the present invention.
  • Fig. 3 is the flow chart of the biological probe preparation method provided by the present invention.
  • Fig. 4 is the general preparation flow chart of customizing sialic acid ligand molecule of general formula I of the present invention according to R 2 ⁇ R 1 ⁇ R 3 group introduction sequence;
  • Fig. 5 is the general preparation flow chart of the custom sialic acid ligand molecule of general formula I of the present invention according to the introduction sequence of R 2 ⁇ R 3 ⁇ R 1 groups.
  • the direct detection method of the new coronavirus provided by the present invention is based on the surface Spike protein of the new coronavirus, and its core principle is to use the customized sialic acid ligand molecule to detect the surface S protein of the new coronavirus.
  • the binding effect was detected by recognizing the binding effect of gold nanorod probe, and the binding effect was characterized by the change of the optical absorption signal exhibited by the LSPR effect of the gold nanorod probe.
  • the corresponding detection principles and detection reagents are described in detail as follows:
  • the S protein on the surface of the new coronavirus particles in the sample can recognize the customized sialic acid ligand molecules bound to the gold nanorod probe. Since the surface of the new coronavirus has abundant S protein molecules, and The surface of the gold nanorods also has a multivalent sialic acid ligand structure, so the two are multivalently combined with each other to form an agglomeration structure. Because the gold nanorods have the LSPR effect, they have optical absorption signals, which can change the surface environment. It is very sensitive.
  • the detection reagent involved in the present invention includes a uniformly dispersed aqueous solution of customized sialic acid ligand molecules-gold nanorod biological probes, and S-Au bonds are passed on the surface of the gold nanorods stabilized by cetyltrimethylammonium bromide (CTAB).
  • CTLAB cetyltrimethylammonium bromide
  • a custom sialic acid ligand molecule is immobilized. By introducing the corresponding group on the sugar ring, the sialic acid ligand molecule can better match the sugar binding domain pocket of the S protein on the surface of the new coronavirus, so that it has high specific recognition for the new coronavirus S protein. , high affinity binding effect, the key composition of the detection reagent is shown in Figure 2.
  • the preparation method of the customized sialic acid ligand molecule-gold nanorod biological probe is shown in FIG. 3 ; the preparation flow chart of the customized sialic acid ligand molecule is shown in FIG. 4 or FIG. 5 .
  • the compound of formula 1-4 (15.2 g, 22.83 mmol) was dissolved in dichloromethane (60 mL), triethylamine (4.61 g, 45.66 mmol) was added, p-toluenesulfonyl chloride (6.52 g, 34.25 mmol) was added in batches, and the mixture was stirred overnight , TLC detected that the reaction was complete, methanol was added, and after stirring for 10 minutes, the solvent was evaporated, the residue was dissolved in dichloromethane, the organic phase was washed with brine, dried over anhydrous sodium sulfate, evaporated to remove the solvent, and the residue was dissolved in N,N- Dimethylformamide (20 mL) was added with sodium azide (3.70 g, 57.08 mmol), and the mixture was stirred in an oil bath at 50° C.
  • the compound of formula 1-6 (8.4 g, 16.41 mmol) was dissolved in dichloromethane (40 mL), triethylamine (3.23 g, 31.91 mmol) was added, p-toluenesulfonyl chloride (4.56 g, 23.93 mmol) was added in batches, and the mixture was stirred overnight , TLC detected that the reaction was complete, methanol was added, and after stirring for 10 minutes, the solvent was evaporated, the residue was dissolved in dichloromethane, the organic phase was washed with brine, dried over anhydrous sodium sulfate, and after concentration, the residue was dissolved in N,N-dimethylmethane Formamide (50 mL), potassium thioacetate (3.54 g, 30.97 mmol) was added, and the mixture was stirred in an oil bath at 50° C.
  • the compound of formula 1-7 (7.6 g, 13.00 mmol) was dissolved in methanol (50 mL), sodium methoxide (0.1 M in MeOH) was added, the pH of the reaction solution was adjusted to 8-10, and stirred at room temperature for 15 minutes. TLC detected that the reaction was complete, and added 732 Form H + cation resin, after stirring for 5 minutes, filter off the resin, and concentrate the filtrate to obtain the compound of formula 1 (6.9 g, 97.8%), ESI-MS m/z calcd for [C 21 H 39 N 2 O 12 S] + (M+H) + :542.60,found:542.59.
  • the compound of formula 2-3 (19.3 g, 25.47 mmol) was dissolved in methanol (40 mL), NaOH aqueous solution (1.5 M, 20 mL) was added dropwise, stirred until the reaction was complete by TLC detection, the reaction solution was neutralized, the solvent was evaporated, and the residue was dissolved in 2M NaOH aqueous solution (40 mL) was refluxed in an oil bath at 95°C for 10 hours, the reaction solution was neutralized, and the solvent was evaporated.
  • the compound of formula 2-5 (18.1 g, 24.4 mmol) was mixed with methyl acrylate (2.46 g, 29.28 mmol) and cuprous iodide (0.93 g, 4.88 mmol) and dissolved in THF (60 mL), and N,N was added dropwise.
  • -Diisopropylethylamine DIPEA (4.73gm 36.6mmol) stirred at room temperature for 1 hour, evaporated the reaction solvent, the residue was dissolved in dichloromethane, successively passed through 1M ammonia water, 1N HCl aq , sat.NaHCO 3a.q.
  • the compound of formula 2-6 (19.6 g, 23.73 mmol) was dissolved in methanol (100 mL), sodium methoxide (0.1 M in MeOH) was added, the pH of the solution was adjusted to 8-10, stirred at room temperature for 30 minutes, and hydrochloric acid was added dropwise to neutralize the reaction solution, The solvent is evaporated to obtain the crude product of formula 2-7, which is directly used in the next step reaction;
  • the compound of formula 2-9 (13.1 g, 17.22 mmol) was dissolved in acetonitrile (50 mL), acetone acetal (5.38 g, 51.66 mmol) and a catalytic amount of p-toluenesulfonic acid (0.3 g, 1.72 mmol) were added, and the mixture was heated at 80° C.
  • the crude compound of formula 2-12 obtained in the previous step was dissolved in dichloromethane (60 mL), triethylamine (2.88 g, 28.47 mmol) was added, and after stirring for 5 minutes, p-toluenesulfonyl chloride (4.07 g, 21.35 mmol) was added in batches at room temperature.
  • the compound of formula 2-13 (8.9 g, 11.98 mmol) was dissolved in methanol (30 mL), an aqueous NaOH solution (1.5 M, 10 mL) was added, stirred for 30 minutes at room temperature, and the reaction solution was neutralized by adding 732 type H + cation resin, and the resin was filtered off. , the filtrate was concentrated to obtain the compound of formula 2 (7.5g, 91.2%), ESI-MS m/z calcd for [C 29 H 43 N 4 O 13 S] + (M+H) + : 687.73, found: 687.72;
  • the preparation process of gold nanorod solution includes: adding chloroauric acid HAuCl 4 aqueous solution (0.5mM, 5 mL) to CTAB aqueous solution (0.2 M, 5 mL), adding freshly prepared sodium borohydride NaBH 4 ice aqueous solution (10 mM) under vigorous stirring , 600 ⁇ L), continue to stir for 10 minutes to prepare gold nanoseed solution; add HAuCl 4 aqueous solution (5 mM, 6 mL), silver nitrate AgNO 3 aqueous solution (0.1 mM, 75 ⁇ L) to CTAB aqueous solution (0.2 M, 30 mL), continue to add Hydrochloric acid (1.2 mM, 75 ⁇ L) and sodium ascorbate Vc-Na aqueous solution (10 mM, 3.7 mL), shake the solution until colorless, add the prepared gold nanoseed solution (50 ⁇ L), stir for 5 minutes, and place in a 37°C incubator Let stand for 16 hours, centrito
  • the preparation process of gold nanorod solution includes: adding chloroauric acid HAuCl 4 aqueous solution (0.5mM, 5 mL) to CTAB aqueous solution (0.2 M, 5 mL), adding freshly prepared sodium borohydride NaBH 4 ice aqueous solution (10 mM) under vigorous stirring , 600 ⁇ L), continue to stir for 10 minutes to prepare gold nanoseed solution; add HAuCl 4 aqueous solution (5 mM, 6 mL), silver nitrate AgNO 3 aqueous solution (0.1 mM, 75 ⁇ L) to CTAB aqueous solution (0.2 M, 30 mL), continue to add Hydrochloric acid (1.2 mM, 75 ⁇ L) and sodium ascorbate Vc-Na aqueous solution (10 mM, 3.7 mL), shake the solution until colorless, add the prepared gold nanoseed solution (50 ⁇ L), stir for 5 minutes, and place in a 37°C incubator Let stand for 16 hours, centrito
  • the new coronavirus particles are highly infectious and highly pathogenic, and the detection of the new coronavirus in the present invention only involves the surface S protein of the new coronavirus, not its internal structure. Therefore, the commercialized new coronavirus S protein can be immobilized on the surface of SiO 2 nanoparticles with a particle size of 100 nm. To simulate the new coronavirus particles, the detection experiments are carried out by simulating the particles.
  • the present invention provides a new coronavirus particle detection method based on the LSPR effect of gold nanorods (Gold nanorods, GNR) probes.
  • Gold nanorods Gold nanorods, GNR
  • Custom sialic acid ligand molecules are immobilized on the surface of gold nanorods, which can efficiently identify and bind to the new coronavirus.
  • the Spike protein on the surface triggers the agglomeration of the dispersed gold nanoprobes, and the change of the dispersion state of the gold nanoprobes can be reflected by the color exhibited by the LSPR effect, thereby realizing the detection of the new coronavirus.

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Abstract

Sont divulgués une sonde biologique, son procédé de préparation et son application. La sonde biologique comprend un support de sonde et une molécule de ligand d'acide sialique personnalisée reliée au support de sonde ; une formule générale développée de la molécule de ligand d'acide sialique personnalisée est telle que représentée dans la formule I ; dans laquelle R1 est choisi parmi un groupe hydroxy, un groupe méthoxy ou un groupe méthoxy substitué ; R2 est choisi parmi un groupe acétamido, un groupe acétamido substitué, un groupe benzoylamino, un groupe benzoyle substitué, un groupe alcoxy carbonyle amide, un groupe triazole ou un groupe triazole substitué ; R3 est choisi parmi un groupe hydroxy, un groupe méthoxy, un groupe méthoxy substitué, un groupe acétamido, un groupe acétamido substitué, un groupe sulfonamide ou un groupe phosphamide ; A est (formule a) ou (formule b), n est choisi parmi 0, 1, 2, 3, 4, 5, 6 ou 7 et m est choisi parmi 2, 3, 4, 5, 6, 7, 8 ou 9. La présente sonde biologique peut être utilisée pour détecter directement une nouvelle particule de coronavirus.
PCT/CN2020/127031 2020-11-06 2020-11-06 Sonde biologique, son procédé de fabrication et son application WO2022094912A1 (fr)

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Cited By (2)

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CN116237532A (zh) * 2022-12-22 2023-06-09 江南大学 一种糖基配体类似物诱导手性钴基超小纳米粒子的合成方法
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