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Monoamine oxidase A microreactor and preparation method and application thereof

A monoamine oxidase and microreactor technology, applied in biochemical equipment and methods, redox enzymes, enzymes, etc., can solve the problems of waste of target enzymes, helpless screening of active compounds, cumbersome operations, etc., achieving high automation and great promotion value , the effect of strong expansion ability

Inactive Publication Date: 2018-12-18
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, spectrophotometry requires a large amount of enzymes and is inefficient, and fluorescence assays also have disadvantages such as high cost and cumbersome operation; moreover, these screening methods usually use free enzymes as test objects, which cause great damage to the target enzymes because they cannot be recycled and reused. waste of
In particular, the above methods can only screen for a single compound, and are helpless for the screening of active compounds in complex systems such as natural products

Method used

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  • Monoamine oxidase A microreactor and preparation method and application thereof
  • Monoamine oxidase A microreactor and preparation method and application thereof
  • Monoamine oxidase A microreactor and preparation method and application thereof

Examples

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Embodiment

[0035] Preparation of the monoamine oxidase A microreactor in this example:

[0036] (1) Take the amino magnetic bead dispersion (Mag NH 2 , 2μm, 10mg / mL, BcMag TM ) 8mg was placed in a 2mL centrifuge tube, solid-liquid separated by a magnetic separator, and the supernatant was discarded to obtain magnetic nanospheres, which were then washed with a pyridine solution (10mM, pH=6.0) for three times and then allowed to stand for use.

[0037] (2) Prepare a 5% (v / v) glutaraldehyde solution with 10mM pyridine solution, mix it with the magnetic nanospheres cleaned in step (1) and place it in a suspension in a chromatography cabinet at 4°C The medium activation reaction is 3 hours, and after the reaction is completed, it is washed three times with a pyridine solution to obtain activated magnetic nanospheres, which are allowed to stand for use.

[0038] (3) Take 20μL of monoamine oxidase A solution (5mg / mL), place it in a 2mL centrifuge tube, add 480μL of pyridine solution (10mM, pH=6.0) a...

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Abstract

The invention belongs to the technical field of drug screening, and discloses a monoamine oxidase A microreactor and a preparation method and application thereof. An amino group-containing magnetic nano material is mixed with a glutaraldehyde solution for activation reaction to obtain an activated magnetic nano material, and the monoamine oxidase A microreactor is prepared by mixing of the activated magnetic nano material and a pyridine solution in which monoamine oxidase A is dissolved for immobilization reaction, performing solid-liquid separation after the immobilization reaction is completed, and removing of supernatant. The monoamine oxidase A microreactor can be applied to screening of antidepressant active ingredients in natural products, has the advantages of simple operation and high automation, and can achieve high-throughput screening.

Description

Technical field [0001] The invention belongs to the technical field of drug screening, and specifically relates to a monoamine oxidase A microreactor and a preparation method and application thereof. Background technique [0002] Monoamine oxidase A (MAOA) is a flavinase that binds to the outer mitochondrial membrane and is widely present in various tissues [H.G. Brunner, et al. American Journal of Human Genetics, 1993, 52, 578.]. In humans, MAOA plays an important role in the oxidative deamination of monoamines in the brain, including neurotransmitters such as serotonin, dopamine, and norepinephrine [SMShelke, et al. Bioorganic&Medicinal Chemistry Letters, 2011, 21 (8):2419-2424.], and detected elevated levels of MAOA in the prefrontal cortex and anterior cingulate cortex in rodent models of human depression and depressive behavior [JHMeyer, et al. Archives of General psychiatry, 2009,66(12):1304-1312; JHMeyer, et al. Archives of General psychiatry, 2010,67(15):468-474.]. MAOA...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12Q1/26
CPCC12N9/0022C12N11/14C12Q1/26C12Y104/03004
Inventor 江正瑾张岩岩张婷婷王启钦陈维佳刘蕊洁刘佳
Owner JINAN UNIVERSITY
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