Optical fiber biosensor and application thereof in homogeneous chemiluminescence biological detection

A homogeneous chemiluminescence and biosensor technology, applied in the field of biochemical analysis, can solve the problems of slow reaction rate, low concentration, hysteresis non-specific signal, etc., and achieve the effects of reduced interference, simple operation, and wide linear range

Active Publication Date: 2021-07-16
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the reaction system, since the concentration of the catalyst in the microenvironment around the luminescent substrate in the immune complex is much higher than that of the solution body, when a sufficient amount of trigger is introduced, the luminescent substrate in the complex reacts rapidly and is consumed, resulting in a Flash signal, while the free luminescent substrate in the solution has a low concentration of catalyst in the surrounding microenvironment, so the reaction rate is slow, resulting in a hysteresis non-specific signal

Method used

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  • Optical fiber biosensor and application thereof in homogeneous chemiluminescence biological detection
  • Optical fiber biosensor and application thereof in homogeneous chemiluminescence biological detection
  • Optical fiber biosensor and application thereof in homogeneous chemiluminescence biological detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Verification of substrate concentration-controlled time resolution in fiber optic sensors

[0060] The biorecognition molecule immobilized on the fiber optic probe was procalcitonin capture antibody, and the chemiluminescence substrate was acridinium ester as an example to verify the substrate concentration-controlled homogeneous chemiluminescence fiber sensor.

[0061] Experiment 1) 10 μg / mL procalcitonin capture antibody and sufficient amount of chemiluminescent substrate acridinium ester-BSA (50 mg / mL) were modified onto the surface of the fiber core to form a fiber optic probe for detecting procalcitonin. Then, the fiber optic probe was reacted with procalcitonin standard substance (10 ng / mL) and HRP-labeled procalcitonin detection antibody to form a sandwich structure, and then a sufficient amount of trigger hydrogen peroxide was added to generate a chemiluminescence signal. Such as figure 1 To record the chemiluminescence kinetic curve. In this chemilu...

Embodiment 2

[0064] Embodiment 2 Construction of fiber optic biosensor

[0065] The main component of the core of the silica fiber is silica, and the biorecognition molecules are fixed on the fiber core through chemical bonds.

[0066] 1. Preparation of fiber optic biosensor

[0067] Taking the biorecognition molecule immobilized by the fiber optic biosensor as procalcitonin capture antibody, and the chemiluminescent substrate as acridinium ester as an example, the preparation process is described. The specific process is as follows:

[0068] 1) Cut the optical fiber into a small section of 6cm, and gently scrape off the acrylic protective layer with a length of 0.5cm at one end of the optical fiber with a knife;

[0069] 2) Corrode the part where the acrylic protective layer is removed with 24% hydrofluoric acid for 20 minutes to remove the silicon dioxide coating layer, rinse with 0.1mol / L NaOH and water three times, and blow dry with nitrogen;

[0070] 3) The fiber core part from whic...

Embodiment 3

[0083] The detection of procalcitonin in embodiment 3 blood

[0084] The principle of procalcitonin detection is as follows: firstly, the conjugate of chemiluminescent substrate acridinium ester and BSA and procalcitonin capture antibody are respectively modified on the optical fiber to prepare a chemiluminescent optical fiber biosensor for detecting procalcitonin. Then the sensor reacts with the test sample and HRP-labeled procalcitonin detection antibody, and an immune sandwich reaction occurs between the capture antibody, the test substance and the detection antibody on the sensor to form a complex, and the HRP in the complex and The concentration of the sample to be tested is related. Then a sufficient amount of trigger hydrogen peroxide is introduced, and the HRP bound on the sensor catalyzes hydrogen peroxide to produce an oxidative product, which immediately reacts with the acridinium ester modified on the optical fiber to generate a luminescent signal, and the intensit...

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Abstract

The invention discloses an optical fiber biosensor, the surface of the optical fiber sensor is simultaneously modified with biological recognition molecules and a chemiluminescence substrate, and the optical fiber sensor can be applied to homogeneous chemiluminescence biological detection. The invention also discloses a homogeneous chemiluminescence biological detection method, in the method, the optical fiber biosensor is used, the chemiluminescence substrate concentration and reaction speed are controlled at the same time, and specific and non-specific detection signals are distinguished from the generation time of a luminescence signal, so that the interference of the non-specific signal is avoided; and the accuracy and the sensitivity of homogeneous detection are improved.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, relates to an optical fiber biosensor, and also relates to the application of the sensor in homogeneous chemiluminescence biodetection. Background technique [0002] Chemiluminescence has high sensitivity and wide linear range, and is widely used in the field of analysis and testing. In particular, chemiluminescent biosensors have both high sensitivity of chemiluminescence and high specificity of biorecognition reactions, and are especially suitable for quantitative analysis of trace targets in complex samples. Compared with other optical biosensors, the equipment is simple and does not require excitation Light source, spectrometer and other components effectively reduce the complexity and cost of the instrument. [0003] Homogeneous determination is an analysis method that does not require separation. The luminescent signal is directly measured after the reaction. This method not only reduce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N21/01G01N33/58
CPCG01N21/76G01N21/01G01N33/581
Inventor 陈翊平何慧禹聂荣彬
Owner HUAZHONG AGRI UNIV
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