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A kind of sugar chain polymer modified microsphere material and preparation method and application thereof

A technology of polymers and microspheres, which is applied in the analysis of materials, material separation, instruments, etc., can solve the problems of cumbersome steps, time-consuming, virus inactivation, etc., and achieve the advantages of easy enrichment or separation, effective capture of viruses, and avoiding hydrolysis Effect

Active Publication Date: 2022-07-19
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional virus separation and purification techniques include ultracentrifugation, ultrafiltration membranes, and various ion-exchange chromatography. Not only are the steps cumbersome and time-consuming, but they are also likely to cause virus inactivation and reduce virus recovery. At the same time, it is difficult for the above-mentioned techniques to achieve automated closed Continuous production, there is a risk of infection for production technicians

Method used

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  • A kind of sugar chain polymer modified microsphere material and preparation method and application thereof
  • A kind of sugar chain polymer modified microsphere material and preparation method and application thereof
  • A kind of sugar chain polymer modified microsphere material and preparation method and application thereof

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preparation example Construction

[0076] The present invention provides the preparation method of the sugar chain polymer modified microsphere material according to the technical solution, comprising the following steps:

[0077] Acrylamide, nitrogen-(4-trimethylsilyl-3-butyne) acrylamide and chain transfer agent are mixed, and RAFT polymerization is carried out under the action of an initiator to obtain acrylamide copolymer with silane-protected side chain alkynyl groups The chain transfer agent is an end-group biotin-functionalized trithioester chain transfer agent or an end-group desthiobiotin-functionalized trithioester chain transfer agent;

[0078] performing deprotection reaction on the acrylamide copolymer of the silane-protected side chain alkynyl group to obtain the acrylamide copolymer of the end group biotinylated side chain alkynyl group or the acrylamide copolymer of the dethiobiotinylated side chain alkynyl group;

[0079] The acrylamide copolymer of the end-group biotinylated side-chain alkynyl...

Embodiment 1

[0137] Non-natural sialic acid oligosaccharide S-6'-sialyllactose (ie, corresponding to the compound numbered No. 3 in Table 2) according to figure 1 The synthetic route shown is prepared, including the following steps:

[0138] Add acidic resin to the methanol solution (3 g / 100 mL) of sialic acid (compound 1) to control the pH value of the system at 3-5, stir at room temperature (25°C) until all dissolved, continue stirring overnight, and remove the acidic resin by filtration , the obtained filtrate was concentrated to obtain compound 2;

[0139] In an ice-water bath (0°C), compound 2 (2.0g), acetyl chloride (20mL), methanol (3mL) and acetic acid (10mL) were added to the pressure vessel, and the reaction was sealed and stirred for 36h at room temperature; the resulting product system was Concentrate to obtain compound 3;

[0140] The obtained compound 3 was dissolved in dry dichloromethane (30 mL), then potassium thioacetate (4 g) was added, and the mixture was stirred over...

Embodiment 2

[0148] Reducing terminal azide sialic acid oligosaccharide was prepared based on sialyllactose (specifically, the compound No. 1 in Table 2, the compound No. 5 or the compound No. 3 prepared in Example 1) was prepared , including the following steps:

[0149] Sialyl lactose (0.65 g), 2-chloro-1,3-dimethylimidazolinium chloride (DMC, 0.52 g), triethylamine (2.0 mL) and sodium azide (0.67 g) were mixed, Under the condition of ice-water bath (0°C), the azide reaction of the water-phase end group was carried out for 1.5 h to introduce the azide group at the reducing end of sialyllactose; the obtained product system was concentrated, and the obtained residue was mixed with N,N- Dimethylformamide (20 mL) was mixed, and unreacted sodium azide was removed by filtration; the filtrate was concentrated, the obtained residue was dissolved in water (20 mL), the obtained system was washed with dichloromethane, and the aqueous phase was collected, Purified by cation exchange resin (IR-120) ...

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Abstract

The invention provides a sugar chain polymer modified microsphere material and a preparation method and application thereof, belonging to the technical field of biological materials. The present invention utilizes the sugar cluster effect of the sugar chain polymer to realize the specific capture of the virus or virus-like particles in the sample, and the sugar chain polymer is modified on the surface of the modified microsphere, so as to facilitate the rapid separation of the virus or virus-like particles from the sample purification. Using the sugar chain polymer-modified microsphere material, the target virus or virus-like particles can be rapidly enriched or purified by pre-processing mixed samples containing viruses or virus-like particles, thereby effectively improving the performance of existing virus detection methods. Sensitivity and accelerate research and development of viral and viral vector vaccines. At the same time, when the microspheres in the sugar chain polymer modified microsphere material are agarose gel microspheres or silica gel microspheres, the present invention also provides its application as a virus affinity chromatography column filler, which can realize the target virus in the virus culture solution. or one-step sequential rapid purification of virus-like particles.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a sugar chain polymer modified microsphere material and a preparation method and application thereof. Background technique [0002] Virus nucleic acid detection requires expensive equipment and a long time. At present, the colloidal gold kit for influenza virus is a rapid detection method widely used in clinical practice. However, affected by the sampling operation and the severity of the disease, the false-negative results caused by the lower virus concentration in throat swab samples have always been a major problem that plagued clinical diagnosis and affected patient treatment. [0003] In addition, in the actual production process of virus vaccine, in addition to the desired virus, the virus medium also contains impurities such as nutrients and proteins and genetic materials from cultured cells, which are all potential allergens. Therefore, in the preparation of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/60C08F230/08C08F220/56C08F8/00C08F8/48
CPCG01N30/60C08F230/085C08F8/48C08F8/00C08F220/56
Inventor 叶新山李格非
Owner PEKING UNIV
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