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Zanamivir-magnetic nanoparticle conjugate, its preparation method and use

A technology of magnetic nanoparticles and zanamivir, applied in the field of zanamivir-magnetic nanoparticle conjugates and their preparation, can solve the problem of high cost of antibody-magnetic nanoparticles, long period of antibody preparation, and expensive antibody, etc. problems, to achieve the effect of being conducive to preservation and application, simple and convenient preparation, and high capture rate

Active Publication Date: 2021-06-08
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because antibodies are expensive and easily denatured, the prepared antibody-magnetic nanoparticles are expensive and difficult to store, even if they are recovered, they are difficult to reuse
On the other hand, due to the high specificity of antibodies, even different strains of the same subtype of influenza virus need to be separately enriched and isolated
For new mutant strains of unknown subtypes, the applicability of the above method has great limitations due to the long production cycle of targeted antibodies.

Method used

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  • Zanamivir-magnetic nanoparticle conjugate, its preparation method and use
  • Zanamivir-magnetic nanoparticle conjugate, its preparation method and use
  • Zanamivir-magnetic nanoparticle conjugate, its preparation method and use

Examples

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Embodiment

[0207] Unless otherwise specified, the methods used in the present invention below are conventional methods; the materials and reagents used can be obtained from commercial sources.

[0208] Reagents used below

[0209] Tris-HCl buffer solution (100 mM, pH 8.0): Tris was dissolved in water at a final concentration of 12.1 g / L, and the pH was adjusted to 8.0 with HCl.

[0210] PBS buffer (10 mM, pH 7.4): NaH 2 PO 4 0.24g / L, Na 2 HPO 4 1.42g / L, KCl 0.2g / L and NaCl 8.0g / L, the solvent is water.

[0211] PBST buffer (10 mM, pH 7.4): NaH 2 PO 4 0.24g / L, Na 2 HPO 4 1.42g / L, KCl 0.2g / L, NaCl 8.0g / L and Tween-20 0.5ml / L, the solvent is water.

[0212] TBS buffer (10mM, pH 7.4): Tris 3g / L, KCl 0.2g / L and NaCl 8.0g / L, solvent is water.

[0213] TBST buffer (10mM, pH 7.4): Tris 3g / L, KCl 0.2g / L, NaCl 8.0g / L and Tween-200.5ml / L, solvent is water.

[0214] 2-(4-morpholine)ethanesulfonic acid (MES, 50nM, pH 5.0) activation buffer, MES dissolved in water, the final concentra...

preparation example 1

[0221] Preparation Example 1 N 3 CH 2 (CH 2 OCH 2 ) 5 CH 2 NH 2 Synthesis

[0222] The compound synthesized in this preparation example is N 3 CH 2 (CH 2 OCH 2 ) 5 CH 2 NH 2 (i.e. the compound shown in formula X-3, m=5), its synthetic route is as follows:

[0223]

[0224] The synthesis of compound 1: Hexapolyethylene glycol (1.41g, 5mmol) was dissolved in triethylamine (3.5mL, 25mmol) and dry dichloromethane (100mL), ice-bathed, p-toluenesulfonyl chloride (2.4 g, 15 mmol) was stirred at room temperature for 24 hours. TLC (EtOAc) detected the completion of the reaction. Add dichloromethane (200mL) to dilute, successively wash with 1M HCl﹑saturated NaHCO 3 and saturated NaCl extraction, with anhydrous NaCl 2 SO 4 The organic phase is dried. Filter to remove anhydrous Na 2 SO 4 Afterwards, the solvent of the organic phase was evaporated to dryness under reduced pressure. The resulting mixture was separated and purified by silica gel column chromatography...

preparation example 2

[0227] Preparation Example 2 Synthesis of Zanamivir Modified with Alkyne Group

[0228] The zanamivir modified with alkynyl group synthesized in this preparation example is compound 5, (i.e. in the compound shown in formula X-2, m=6), and its synthesis route is as follows:

[0229]

[0230] Compound 4 was prepared according to the method disclosed in CN103819665B. 156mg of compound 4 (0.167mmol) was put into a 25mL reaction flask, dissolved in 10ml of methanol, the pH of the reaction solution was adjusted to 13 with 1M NaOH aqueous solution, and stirred at room temperature for 3 hours. TLC monitors reaction process, adds acidic resin Dowex 50 (H + ) to neutralize and adjust the pH of the reaction solution to about 7. After being filtered and spin-dried, the obtained crude product was dissolved in a mixed solution of trifluoroacetic acid and dichloromethane (trifluoroacetic acid:dichloromethane=1:1), and stirred at room temperature for 3 hours. The progress of the reactio...

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Abstract

The present invention relates to a zanamivir-magnetic nano particle conjugate, its preparation method and its use in detecting influenza virus, NA protein and RNA of the virus. The zanamivir-magnetic nanoparticle conjugate of the present invention can enrich, separate and / or purify almost all types of influenza viruses and / or NA proteins in samples with low concentration and complex components, so that the influenza virus , The extraction and / or detection of the NA protein or RNA of the virus is simpler and faster.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to a zanamivir-magnetic nanoparticle conjugate and a preparation method thereof, and the use of the conjugate in detecting influenza virus, NA protein or RNA of influenza virus. Background technique [0002] Influenza (flu) is a zoonotic infectious disease caused by influenza virus, and its host involves animals such as people, pigs, birds, horses and dolphins. Influenza viruses mutate quickly, have high transmissibility, and are highly harmful. In addition to causing seasonal influenza, each large-scale epidemic of influenza virus has caused great losses to human society. For example, the 1918 Spanish flu killed nearly 20 million people, the 1957 Asian flu and the 1968 Hong Kong flu killed 2 million and 1.5 million, respectively. In recent years, due to the emergence of new mutant strains of viruses and drug resistance, cross-species transmission between humans and animals, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/34C12Q1/04C12R1/93
CPCC12Q1/04C12Q1/34C12Q1/6806C12Q1/701G01N2333/11
Inventor 李学兵张振兴
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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