1,3-beta-D glucan derivative, kit, preparation method and method for determining content of 1,3-beta-D glucan

A technology for dextran and derivatives, applied in the fields of 1,3-β-D glucan derivatives and their preparation, kits for measuring 1,3-β-D glucan content and their preparations, reaching High specificity, improvement of low sensitivity, and simplified experimental procedures

Inactive Publication Date: 2021-04-09
昆明思安生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problems in the background technology, the existing 1,3-β-D glucan kits cannot be used for both solid-phase coupling and luminescence-labeled 1,3-β-D glucan Defects of derivatives, the biggest feature of this technical solution is to provide a 1,3-β-D glucan antigen derivative and its preparation method, through the preparation of derivatives, 1,3-β -D glucan antigen is indirectly connected with biotin or enzyme, which solves the key problem of the development of this project, that is, the coupling problem, and can conveniently establish a magnetic particle enzymatic chemiluminescent system of 1,3-β-D glucan

Method used

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  • 1,3-beta-D glucan derivative, kit, preparation method and method for determining content of 1,3-beta-D glucan

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Embodiment

[0044] A 1,3-β-D glucan antigen derivative, the preparation steps of the 1,3-β-D glucan antigen derivative are:

[0045] 1) Prepare 0.1M HCl solution;

[0046] 2) Prepare 0.01M of PH=9.0 and 0.15M of PH=9.5 respectively NaHCO buffer;

[0047] 3) Weigh about 10 mg of 1,3-β-D glucan and dissolve it to 10 mg / mL with N,N-dimethylformamide (DMF) buffer;

[0048] 4) Weigh 50mg of succinic anhydride and dissolve in 10ml DMF to 5mg / mL;

[0049] 5) Weigh 20 mg of lysine and dissolve it in 1 mL of 0.01M NaHCO3 buffer solution with pH=9.0 to 20 mg / mL;

[0050] 6) Add 0.5mL of 1,3-β-D glucan-DMF solution to 10mL of succinic anhydride-DMF solution, and react at 65°C for 6h;

[0051] 7) After the reaction is completed, add 1.2mL tributylamine and 0.65mL isobutyl chloroformate, and react at room temperature for 30min;

[0052] 8) Add 1 mL of lysine solution with a concentration of 20 mg / mL to the previously reacted 1,3-β-D glucan-succinic anhydride in DMF solution, and react at room temp...

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Abstract

The invention discloses a 1,3-beta-D glucan derivative, a kit, a preparation method and a method for determining the content of 1,3-beta-D glucan. The invention provides a 1,3-beta-D glucan magnetic particle chemiluminiscence immunoassay kit which combines a magnetic particle separation technology, a biotin-avidin amplification technology and a chemiluminiscence analysis technology. Meanwhile, a biotinylated 1,3-beta-D glucan antigen derivative and an enzyme-labeled 1,3-beta-D glucan antigen derivative are prepared from the 1,3-beta-D glucan antigen derivative, so that two systems (1, the biotinylated 1,3-beta-D glucan antigen derivative plus an antibody enzyme-labeled reagent and plus a calibration solution; 2, the enzyme-labeled 1,3-beta-D glucan antigen derivative plus a biotinylated 1,3-beta-D glucan mouse monoclonal antibody and plus calibration liquid) are constructed, and the 1,3-beta-D glucan in a sample can be tested, so that different enterprises have more flexible selection when developing the kit, and the kit can be realized by only preparing the intermediate, namely, the 1,3-beta-D glucan antigen derivative.

Description

technical field [0001] The invention belongs to the field of biochemical detection, and more specifically relates to a 1,3-beta-D glucan derivative and a preparation method thereof and the determination of the content of 1,3-beta-D glucan by using magnetic particle chemiluminescence method Kit and preparation method thereof. Background technique [0002] 1,3-β-D glucan is a kind of polysaccharide, widely exists in fungal cell walls except zygomycetes, accounts for more than 50% of fungal cell wall components, and is a unique component on fungal cell walls. After the fungus enters the blood or deep tissues of the human body, 1,3-β-D glucan can be released from the cell wall through phagocytosis and digestion by phagocytic cells, increasing the content in blood and other body fluids; while in superficial fungal infections , 1,3-β-D glucan is not released, so its content in body fluids does not increase, and the detection of 1,3-β-D glucan in blood or non-blood samples can be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/02G01N21/76G01N33/53G01N33/535G01N33/543G01N33/577
CPCC08B37/0024G01N33/577G01N33/54326G01N33/535G01N33/5308G01N21/76G01N2400/24
Inventor 鲁衡高燕孙金超石云齐
Owner 昆明思安生物科技有限公司
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