Capillary enzyme array micro-reactor and its application

A micro-reactor and capillary technology, applied in instruments, scientific instruments, measuring devices, etc., can solve the problems of difficult miniaturization, complicated manufacturing steps, and limited applications.

Inactive Publication Date: 2013-05-08
MINZU UNIVERSITY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The capillary electrophoresis enzyme microreactors currently researched and reported are essentially single-type enzyme microreactors, and the manufacturing steps are relatively complicated.
And this type of reactor can only be studied for the biochemical behavior of one enzyme, which limits its application and is not suitable for high-throughput research
Although capillary electrophoresis enzyme microreactor arrays have been reported in the prior art, this array requires the application of multiple capillaries, making it difficult to miniaturize

Method used

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  • Capillary enzyme array micro-reactor and its application
  • Capillary enzyme array micro-reactor and its application
  • Capillary enzyme array micro-reactor and its application

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0019] The specific preparation process of the capillary enzyme array microreactor: install a 50 cm long bare capillary on a capillary electrophoresis apparatus, pretreat with 0.1 mol / L hydrochloric acid for 30 minutes, and rinse with deionized water for 10 minutes. Inject the 3-aminopropyltrimethoxysilane ethanol solution (10%) with a length of 41.78mm at the injection end of the capillary with a pressure of 0.5psi×65s, and incubate the reaction for 120min. Reverse air for 10 minutes to remove unbonded silylating reagents, and immerse the sample inlet and sample outlet in deionized water for 5 minutes for cleaning. After the derivatization of the amino group is completed, inject a 9.64mm section of Trypsin solution (4u / ml) with an injection pressure of 0.5psi×15s. Air injection was used to push the enzyme solution to a position 32.14 mm away from the capillary injection port, and after 10 minutes of reaction, the air was reversed to remove unbonded Trypsin. Inject a 9.64mm m...

Embodiment 2

[0023] Using the capillary enzyme array microreactor of Example 1, three kinds of enzyme microreactors were constructed on the inner wall of the quartz capillary (50 μm in inner diameter, 40 cm in effective length), specifically, the inlet port has a PNP microreactor with a length of 9.64 mm; There is an EGFR enzyme microreactor with a length of 9.64mm at the position of 16.07mm from the end; there is a Trypsin microreactor with a length of 9.64mm at the position of 32.14mm from the inlet end. The array reactor was installed on a capillary electrophoresis apparatus. Based on the capillary electrophoresis procedure, H 2 O, 100mmol / L borax-boric acid buffer solution pH 8.0, followed by pressure washing, injecting 1mmol / LBAEE into the position of the array microreactor under pressure, the length does not exceed the total length of the enzyme array area, after incubation for 5min, apply 25kV forward voltage to separate. Use ultraviolet detection to collect the electrophoresis sp...

Embodiment 3

[0026] The capillary enzyme array microreactor prepared in Example 1 was used and installed on a capillary electrophoresis instrument. Based on the capillary electrophoresis procedure, H 2 O, 100mmol / L borax-boric acid buffer solution pH 9.5 was pressure-rinsed in sequence, and the mixed solution of 1mmol / LBAEE, ATP and inosine was injected into the reactor under pressure, and the length covered the enzyme array area. After incubation for 5 minutes, apply 25kV forward voltage to separate. Use ultraviolet detection to collect the electrophoresis spectrum at 200nm, and the ultraviolet absorption peaks of the substrate and product can be obtained. image 3(From left to right: 1. BAEE, 2. ATP, 3. Hypoxanthine, 4. Inosine, 5. BA, 6. ADP).

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Abstract

The invention discloses a capillary enzyme array micro-reactor and its application. The enzyme array micro-reactor is formed through constructing a plurality of enzymes in a capillary in addressable section based on a covalent fixing technology in the invention. The capillary enzyme array micro-reactor integrates a plurality of types of enzyme reactors, and can simultaneously realize the interacting high-flux analysis of the plurality of enzymes within a unit time, so the research period is substantially shortened. The capillary enzyme array micro-reactor has the advantages of simple method, low cost, high analysis efficiency, and easy automation, and has great scientific research values and economic benefits in the high-flux screening and the like of enzyme inhibitors and medicines.

Description

technical field [0001] The invention belongs to the technical field of biochemical microreactor production, relates to capillary electrophoresis, enzyme immobilization, and array enzyme immobilization technologies, and belongs to a novel miniaturized biochemical reaction device of capillary electrophoresis technology. Background technique [0002] Enzyme microreactor is a new type of biochemical reaction device that combines biomolecular immobilization technology with modern biochemical microanalysis methods. This miniaturized reaction system plays a very important role in the fields of life science research such as proteomics and protease inhibitor screening due to the advantages of specific recognition of immobilized enzymes, reusability and low consumption of microanalysis. . Currently the most common technology is the capillary electrophoresis enzymatic microreactor. This technique combines the advantages of specific enzyme recognition and efficient separation by capil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/453
Inventor 申刚义于婉婷张爱芹崔箭曾鸣乌日图
Owner MINZU UNIVERSITY OF CHINA
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