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Preparation of reagent-free ampoul immuno sensor and use thereof

A technology of immunosensors and reagents, applied in instruments, scientific instruments, biological testing, etc., can solve the problems of large sample consumption, cumbersome operation steps, and long analysis time, so as to simplify the operation steps, simplify the analysis system, and shorten the analysis time Effect

Inactive Publication Date: 2003-08-27
JIANGSU CANCER HOSPITAL +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] It can be seen that the conventional immunoassay method has cumbersome operation steps, long analysis time, large sample consumption, and high measurement cost, while the existing amperometric immunosensor needs to add non-immune reagents such as media or substrates to the solution to be tested. Even if the oxygen dissolved in the solution to be tested is used as the medium, the measurement results are also affected by changes in ambient temperature, pressure, solution composition, etc.

Method used

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  • Preparation of reagent-free ampoul immuno sensor and use thereof
  • Preparation of reagent-free ampoul immuno sensor and use thereof

Examples

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Embodiment 1

[0034] 1. Example 1: Preparation of CA125 immunosensor

[0035] Choose a disk-shaped planar glassy carbon electrode with a diameter of 4mm as the carrier material, and take CA125 as an example for embedding and fixing:

[0036] (a) Electrode polishing: Firstly, the surface of the electrode is polished with metallographic sandpaper, and then polished on the suede with 1.0, 0.3, and 0.05 μm γ-alumina slurry respectively, and then rinsed with secondary water, and the surface of the electrode is washed with 1 : 1 Ultrasonic cleaning with nitric acid, acetone, and secondary water to obtain a fresh and clean electrode surface.

[0037] (b) Electrode indication pretreatment: Electrolyze the polished electrode in a phosphate buffer solution with a pH value of 5.0 at a constant potential of +1.75V for 300 seconds, and then test it at +0.3V~+1.3V and +0.3V respectively Circular scanning within the potential range of ~-1.3V until a stable current value is obtained. The electrodes were ...

Embodiment 2

[0040] 2. Embodiment 2: Determination of CA125

[0041] (a) Optimization of measurement conditions: respectively change the incubation temperature, incubation time and the amount of HRP-labeled CA125 antibody in the reaction solution, and select the corresponding amount at the time of the maximum current response as the optimal measurement condition. The maximum current response was obtained by incubating at high temperature for 40 min. When the amount of HRP-labeled CA125 antibody in 50 microliters of the reaction solution was greater than 83%, the current response reached the maximum value.

[0042] (b) Determination of CA125: Under optimized experimental conditions, a series of standard CA125 antigen concentrations were measured by competitive immunoassay method to determine the standard curve for CA125 determination, and then the standard curve method was used to determine the concentration of CA125 antigen in the sample.

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Abstract

The antigen to be tested is dissolved in the buffer solution. After the pretreatments of carrier surface, said solution is dropped and coated on said surface which is then over alkoxy compound. The immunity sensor can be obtained by sealing the aqueous thermostat of the system. With the condition of measuring antigen to be tested being opimized, the standard curve for measuring the antigen to be tested can be obtained after a series of treatments are carried out. Through the degradation of the electrochemical signal, the correspondent concentration can be looked up from the standard curve. The method does not need to add the additional intermediate or reagent into the solution of the specimen to be measured so as to prevent the pollution of the electrodes as well as reduce the operation steps.

Description

1. Technical field [0001] The invention relates to the preparation and application of an immune sensor, in particular to the preparation of a reagent-free amperometric type immune sensor and its application in immune analysis. 2. Background technology [0002] In recent decades, immunoassay technology has combined the specific recognition reaction between antigens and antibody molecules and the sensitivity and convenience of electrochemical, spectroscopy, acoustics and other technologies, with its high selectivity for analytes such as tumor markers. And high sensitivity, it has become one of the important analytical methods in clinical, biochemical, environmental analysis and other fields. Conventional immunoassay methods mainly include: radioimmunoassay, enzyme-linked immunoassay, fluorescence immunoassay, chemiluminescence immunoassay, flow injection immunoassay, etc. The above method mainly utilizes the labeling of radioactive isotopes, enzyme molecules, fluorescent subs...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N33/53G01N33/535
Inventor 严枫戴宗鞠熀先
Owner JIANGSU CANCER HOSPITAL
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