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Serum sugar spectrum parting method based on micro-fluidic chip

A technology of microfluidic chip and typing method, which is applied in the field of sugar analysis, can solve the problems of long-term separation mode, etc., and achieve the effect of rapid separation and analysis ability, high-efficiency separation and detection, and high-resolution

Active Publication Date: 2017-06-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

However, this method still requires a long time and different separation modes for glycotyping.

Method used

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  • Serum sugar spectrum parting method based on micro-fluidic chip
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  • Serum sugar spectrum parting method based on micro-fluidic chip

Examples

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Embodiment 1

[0018] 1) Selection of electrophoresis buffer system:

[0019] The injection buffer system consists of 50mmol / L tris(Tris), 50mmol / L N-tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid (TAPS), 1mmmol / L ethylenediaminetetra Acetic acid (EDTA) is composed of 1×TTE buffer solution, pH8.3; the sample separation system is composed of POP 4 or POP 7 gel;

[0020] 2) Preparation and labeling of serum samples

[0021] Take 100μL of blood, centrifuge at 3000r / min for 10min to separate the serum, add 2μL of 10mmol / l NH 4 HCO 3 Buffer and 3 μL deionized water, heat at 95°C for 5 minutes, put in the refrigerator (4°C) for about 15 minutes, add N-glycopeptidase F, incubate in a 37°C incubator for 12 hours (or overnight), and put it in the refrigerator (4°C) ) in an ice bath for 15 minutes, centrifuged for 15 minutes, and the supernatant after centrifugation was used for determination; take 6 μL of centrifuged and dried at 60°C for 120 minutes without a cover (to ensure that the tube...

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Abstract

The invention discloses a serum sugar spectrum parting method based on a micro-fluidic chip, and relates to the field of sugar analysis. The method comprises the steps of selecting a buffer system, selecting an electrophoresis chip, a serum sample, carrying out labeling and electrophoresis, and the like. By combination of a chip electrophoresis technology and a laser-induced fluorescence detection system, 12 N-sugar chains appearing in serum are separated within 8 minutes, so that the shortcoming that a conventional bio-chemical blood analysis method cannot be used to analyze an N-sugar spectrum is overcome. With high resolution and high separation and analysis capacity for the N-sugar spectrum of the serum, chip electrophoresis is expected to be one of analysis measures for quickly, conveniently and efficiently separating and detecting the N-sugar spectrum of the clinical serum sample.

Description

technical field [0001] The invention relates to the field of sugar analysis, in particular to a microfluidic chip-based serum sugar profile typing method. Background technique [0002] All cells and most macromolecules in nature are covered with sugars. Sugars are closely related to life activities. They play an important role of "recognition marks" in the recognition, adhesion and signal transduction between cells, cells and matrix or foreign objects. In recent years, studies have found that there are often changes in sugar chains on the surface of malignant tumor cells, and the relationship between sugar chain structure and diseases, especially tumors, has attracted more and more attention. [0003] At present, reports at home and abroad have analyzed the sugar chain structure from multiple aspects. The main methods are: sugar micro-dot matrix, fluorescent sugar-binding probe, nuclear magnetic resonance, high performance liquid chromatography, time-of-flight mass spectrom...

Claims

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Application Information

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IPC IPC(8): G01N27/447G01N21/64
Inventor 秦建华苏文涛姜雷李艳峰
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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