Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof

A chemiluminescence detection and acridinium ester marker technology, which is applied in the field of life science and medical detection, can solve the problems that the instrument has not been widely used, the analysis speed is slow, and the operation is cumbersome, so as to achieve high detection sensitivity, simple equipment, The effect of extending the range of application

Inactive Publication Date: 2010-05-19
FUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] 2) An antigen or antibody used by CLIA can only detect one analytical object. When there are multiple target objects to be detected in the system, they must be labeled and detected separately. The operation is cumbersome and the analysis speed is slow
However, the detection system they designed can only detect the analyte when the pH of the electrophoresis buffer is 3 Better separation can be achieved only under certain circumstances, so the above-mentioned instruments of Michael A. et al. have not been widely used

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  • Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof
  • Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof
  • Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof

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Embodiment 1

[0027] 1. Detection of 4-(2-hydroxysuccinamidocarbonylethyl)phenyl-10-methylacridine-9-carboxylate fluorosulfonate (AE-NHS):

[0028] 1) Electrophoresis conditions: the total length of the separation capillary is 550 mm, the inner diameter is 75 μm; the running buffer is 5 mmol / L, phosphate buffer solution with pH=8.0; the new capillary is washed with 1 mol / L HCl, 1 mol / L NaOH and water in sequence Activate the surface of the column for 20 minutes, wash with 0.1mol / L NaOH and water for 10 minutes before starting work every day, and then wash with running buffer for 5 minutes. Operating voltage: 15kV. Separation temperature: 20°C. Sampling: electric injection 20s.

[0029] 2) Sample detection: use the electrophoresis buffer described in 1) to prepare concentrations of 1.0×10 -11 mol / L, 1.0×10 -10 mol / L, 1.0×10 -9 mol / L, 1.0×10 -8 mol / L, 1.0×10 -7mol / L, 1.0×10 -6 mol / L, each 1ml of AE-NHS standard solution, according to the electrophoresis conditions described in step 1)...

Embodiment 2

[0031] Qualitative and quantitative determination of alanine and lysine:

[0032] 1) Electrophoresis conditions: the total length of the separation capillary is 550mm, and the inner diameter is 75 μm; the running buffer is 20mmol / L, and the acetate buffer solution of pH=5.8 (wherein adding acetonitrile with a volume fraction of 28% is used as an additive); the new capillary uses 1mol / L Rinse the surface of the activation column for 20 min with L of HCl, 1 mol / L NaOH and water in sequence, and rinse with 0.1 mol / L NaOH and water for 10 min before starting work every day, and then rinse with running buffer for 5 min. Operating voltage: 15kV. Separation temperature: 20°C. Sampling: electric injection 15s.

[0033] 2) Luminescent labeling of alanine and lysine standards:

[0034] Weigh 4.0 nmol of alanine and lysine and 2 μl of 2.0 mmol / L AE-NHS in 98 μl of 0.05 mol / L phosphate buffer solution (pH=8.0) for 10-20 minutes at room temperature in the dark.

[0035] 3) Qualitative ...

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Abstract

The present invention provided a capillary electrophoresis chemiluminescence detection apparatus and method for detecting acridinium ester, acridine sulfonamide and compounds labelled thereby. The apparatus in accordance with the present invention comprises a capillary electrophoresis buffer solution storage container, a high pressure pump, a high voltage power supply, a sample injector access port of separation capillary, an interface unit, a photomultiplier, a weak light detector and a computer, wherein the interface unit comprises acidizing capillary, acidizing reaction liquid injection pump, a chemiluminescence detection cell, a luminescence base solution injection pump. The detection method in accordance with the present invention comprises steps of labelling a compound to be detected; separating the compound to be detected using capillary electrophoresis; acidizing separated effluent liquid out of capillary, detecting the chemiluminescence; analyzing qualitatively and quantificationally. The method in accordance with the present invention is simple and convenient, with a simple apparatus structure, a convenient operation and a low cost, is suitable in electrophoresis buffering systems having a pH<=11. Said method is capable of satisfying what most analysis objects need, and being used for separating and detecting amino acids, polypeptides, proteins, nucleic acids as wellas other acridinium esters, acridine sulfonamides and compounds labelled thereby.

Description

technical field [0001] The invention relates to the fields of life science and medical detection, and more specifically relates to a capillary electrophoresis chemiluminescence instrument and detection method for acridinium esters, acridine sulfonamides and markers thereof. Background technique [0002] The chemiluminescent reaction of acridinium esters or acridine sulfonamides does not require a catalyst, and in the presence of H 2 o 2 It can emit light in the presence of NaOH and has many advantages, especially without a catalytic process and no enhancer, thereby reducing the background luminescence, improving the signal-to-noise ratio, less interference, and high luminous efficiency. For example, the quantum yield of acridine aromatic ester can be as high as 0.05 [MayerA, etc. Angew. Chem. Int. Ed. Engl., 1994, 33(10): 1044]. As luminescent markers of chemiluminescent immunoassay (CLIA), this kind of compound also has other advantages, such as fast and concentrated ligh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N27/447
Inventor 陈国南邱彬郭隆华姜鹰雁
Owner FUZHOU UNIV
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