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Lateral Flow Chromatography System for Quantitative Detection of Alpha-fetoprotein Heterosomes

An alpha-fetoprotein antibody and lateral chromatography technology, which is used in measurement devices, analytical materials, biological tests, etc., can solve the problems of expensive reagents, high technical requirements of the i30 detection system, and increased complexity of operation.

Active Publication Date: 2018-05-11
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, plant lectin affinity immunoelectrophoresis technology and The i30 detection system has high technical requirements, cumbersome operation, and expensive reagents, which limit its clinical application
However, the glycosyl capture spin column increases the complexity of the operation because the sample processing and detection are carried out separately.

Method used

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  • Lateral Flow Chromatography System for Quantitative Detection of Alpha-fetoprotein Heterosomes
  • Lateral Flow Chromatography System for Quantitative Detection of Alpha-fetoprotein Heterosomes
  • Lateral Flow Chromatography System for Quantitative Detection of Alpha-fetoprotein Heterosomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1, lateral flow chromatography system and preparation method of the present invention

[0078] (1) Preparation steps of latex particles (latex MP, Merck)

[0079] 1. Latex MP activation

[0080] Take 1mg of Latex MP, wash with 1mL 25mM MES (pH5.0) buffer 3 times (mix well for 10min each time);

[0081] 5). Prepare 10mg / mL EDC (M=191.7g / mol, C=52mM) solution (EDC is dissolved in cold 25mM MES, pH5.0); (EDC, sigma, 39391)

[0082] 6). Resuspend Latex MP with 440 μL of 25 mM MES (pH 5.0).

[0083] 7). Add 10 μL EDC solution, mix well, shake slowly, 30min, RT;

[0084] 8). Centrifuge the centrifuge tube after the reaction (3000rmp, 5min), remove the supernatant, and wash 3 times with 1mL 25mM MES, pH5.0;

[0085] 2. Latex MP coating

[0086] 6). Resuspend lmgLatex MP with 300 μL 25mM MES, pH 5.0;

[0087] 7). Add 5-12ul (preferably 10ul) 5mg / ml LCA (25mM MES, pH5.0) solution;

[0088] 8). Use 25mM MES, pH 5.0 to make the reaction system to a volume of 500μ...

Embodiment 2

[0121] Embodiment 2, sample pad immobilization MP-LCA coating concentration selection

[0122] 1. Preparation of chromatography test strips

[0123] In this embodiment, the lateral flow chromatography system of the present invention is a chromatography test strip, and the preparation method of the sample pad is the same as that of the sample pad in step (3) of Example 1, wherein different concentrations of MP- LCA (0, 0.1, 0.5, 2.5, 10, 50ug / cm 2 ) (calculated based on LCA concentration) was applied to the sample pad.

[0124] 2. Detection of AFP-L3 binding efficiency under different concentrations of MP-LCA

[0125] 1) Add 100 μL serum containing different concentrations of AFP-L3 (2, 8, 40, 160, 400ng / ml) (the AFP concentration is known), RT, 15min;

[0126] 2) Then calculate the concentration of AFP-L3 with a strip reader, according to the binding ability of lectin to AFP-L3 of different concentrations [(actual measured concentration / theoretical concentration)×100%.

[...

Embodiment 3

[0128] Embodiment 3, using the lateral flow chromatography system of the present invention to carry out clinical sample detection

[0129] The method of using the lateral flow chromatography system

[0130]

[0131] The specific operation is:

[0132] Take the chromatographic kit described in Example 1 or the chromatographic test strip described in Example 2, drop 100 μL of the serum to be tested at the sampling point, and wait at room temperature for 15 minutes;

[0133] Then read the concentration value on the strip reader (C10066-10, Hamamatsu), and calculate the AFP-L3 concentration in the sample according to the measured sample AFP concentration (that is, the AFP-L3 concentration=sample AFP concentration-chromatographic test strip reading value ).

[0134] A total of 324 clinical samples were collected, including 103 HCC (primary liver cancer) samples, 63 non-HCC (chronic hepatitis, cirrhosis) samples, and 46 normal human serum samples.

[0135] Adopt the method of ...

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Abstract

The invention provides a lateral chromatography system for quantificationally detecting alpha-fetoprotein variants, and relates to the field of medical reagents. The lateral chromatography system sequentially comprises a sample region and a combining region in the chromatography direction, wherein agglutinin activated emulsion particles coat the sample region, and are used for combining alpha-fetoprotein variants in a liquid sample; colloidal gold combining articles coat the combing region and are used for combining alpha-fetoprotein in the liquid sample; the coating concentration of the agglutinin activated emulsion particles on the sample region is 2.5 to 50 ug / cm<2>; the agglutinin is lens culinaris agglutinin. The lateral chromatography system has the advantages that the operation is simple; convenience and high speed are realized; the cost is low; the SFP-L3 separation and determination are realized in one step; the efficiency is higher.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations, and in particular relates to a lateral flow chromatography system for quantitative detection of alpha-fetoprotein heterogeneity. Background technique [0002] It is generally believed that alpha-fetoprotein (AFP) is a relatively specific tumor marker for HCC (hepatocellular carcinoma, hepatocellular carcinoma), and sustained elevation of AFP is a risk factor for HCC. At present, some European and American scholars believe that the sensitivity and specificity of AFP are not high. The 2010 edition of the American Society for the Study of Liver Diseases (AASLD) guidelines no longer use AFP as a screening indicator, but most HCCs in my country are related to HBV (hepatitis B virus) infection. Different from the pathogenic factors of HCC in western countries (mostly HCV, alcohol and metabolic factors), combined with the results of domestic randomized research (RCT) and the actual situation, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57438G01N33/57484G01N33/68G01N33/6893G01N2333/471G01N2800/085
Inventor 肖智李全焦守恕
Owner 同昕生物技术(北京)有限公司
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