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Immunosensor based on photonic crystal fiber and application thereof

A technology of photonic crystal fiber and immune sensor, which is applied in the direction of instruments, scientific instruments, material inspection products, etc., and can solve problems such as detection sensitivity limitations

Active Publication Date: 2017-01-04
NORTHEAST NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the above advantages, the capillary is a one-dimensional cylinder, and the detection sensitivity is limited to a certain extent.

Method used

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  • Immunosensor based on photonic crystal fiber and application thereof
  • Immunosensor based on photonic crystal fiber and application thereof
  • Immunosensor based on photonic crystal fiber and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Sources of reagents and equipment used:

[0040] 1 Alpha-fetoprotein antigen and its antibody: Beijing Boaosen Biotechnology Co., Ltd.;

[0041] 2 Antigen diluent: Beijing Keyue Zhongkai Biotechnology Co., Ltd.;

[0042] 3 Oxidized Glutathione: Sigma Aldrich;

[0043] 4 Aminopropyldiethoxymethylsilane, Glutaraldehyde: Sigma Aldrich;

[0044]5 LMA-20 photonic crystal fiber: NKT Photonics (Denmark)

[0045] 6 Diode lasers: Cobolt AB, (Sweden)

[0046] 7 Optical components: Beijing Zhuoli Hanguang Instrument Co., Ltd.

[0047] 8 Photomultiplier tube: Beijing Hamamatsu Photonics Trading (China) Co., Ltd.

[0048] 9 Chromatographic Workstation: Shanghai Vientiane Instrument Co., Ltd.

[0049] Fabrication of photonic crystal fiber immunosensor

[0050] 1) Pretreatment of photonic crystal fiber

[0051] Wash the PCF with 0.1M HCl and 0.1M NaOH successively for 30 min and 60 min respectively to activate the silanol on the inner surface. After washing with water for 30m...

Embodiment 2

[0057] 1. Preparation of AFPA standard curve

[0058] 1) Use antigen diluent to prepare a series of alpha-fetoprotein antigen standard substances with different concentrations, pass them into different root PCFs sequentially, and perform immunoreaction at 37°C for 60 minutes, and wash away unbound AFP with PBST.

[0059] 2) AF-488-labeled alpha-fetoprotein antibody (Ab 2 ) into PCF, immunoreact at 37°C for 60 minutes, wash off unbound Ab with PBST 2 .

[0060] 3) Use a knife to remove the acrylate coating (about 1 cm in length) on the surface of the PCF, which serves as the LIF detection window.

[0061] 4) Set the laser power to 1mW, and the photomultiplier tube (PMT) collects the fluorescence intensity at 520nm.

[0062] 5) Draw the standard curve with the fluorescence intensity as the vertical axis and the AFP concentration as the horizontal axis. The minimum detection limit of the antigen is 0.1ng / mL, such as Figure 6 shown. The minimum detection limit of the capill...

Embodiment 3

[0073] Actual sample testing

[0074] Take a serum sample, add AFP standard samples of different known concentrations, and calculate the recovery rate of the spiked standard. The results are listed in the table below.

[0075]

[0076] 10 actual serum samples with known concentrations were detected with the sensor prepared by the present invention. Substitute the fluorescence intensity into the standard curve, and compare the result with the known concentration, such as Figure 9 It shows that the method has high accuracy.

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Abstract

The invention belongs to the technical field of immunoassay analysis and particularly relates to an immunosensor based on photonic crystal fibers as well as a manufacturing method and application thereof. The manufacturing method comprises the following steps: combining an alpha-fetoprotein antibody Abl on the PCF inner surface through a covalent bond, adding an antigen and a fluorescent labeled second antibody (Ab2) sequentially, forming a sandwich structure Ab1-AFP-Ab2 through the interaction between the antibody and the antigen, and quantitatively detecting a tumor marker AFP of primary hepatic carcinoma (PHC). Compared with an existing product and the prior art, the photonic crystal optical fiber immunosensor has the benefits that the whole preparation process is simple and suitable for industrial production; the antigen concentration is positively correlated with the fluorescence intensity, so that the alpha-fetoprotein can be qualitatively and quantitatively detected; the operation in the detection process is very simple and convenient, and the needs of higher detection sensitivity can be met, therefore, an important clinical diagnostic significance is realized.

Description

technical field [0001] The invention belongs to the technical field of immune detection and analysis. In particular, it relates to a detection method based on photonic crystal fiber optic immunosensor and alpha-fetoprotein antigen. Background technique [0002] Primary liver cancer (Primary Hepatic Carcinoma PHC) is one of the most common malignant tumors clinically. At present, the incidence of liver cancer in the world is on the rise. According to the "Global Cancer Report 2014" published by the World Health Organization, the number of new cancer cases in China ranks first in the world, and the number of new cases and deaths of liver cancer ranks first in the world. Therefore, the early diagnosis and treatment of primary liver cancer is of great significance. [0003] Alpha-fetoprotein (α-fetoprotein or AFP) is a single multimer peptide bond protein, composed of 590 amino acids, with a molecular weight of about 70kD and a sugar content of 4%. AFP is mainly synthesized ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/531G01N33/57438G01N33/57484G01N33/6893
Inventor 杨丽刘晓霞宋星达田苗苗
Owner NORTHEAST NORMAL UNIVERSITY
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