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Exclusion chromatography detection new method of immune complex

A technology of immune complex and size exclusion chromatography, which can be used in measurement devices, analytical materials, instruments, etc., can solve the problems of inability to detect detection objects and low sensitivity, shorten detection time, wide application range, and save washing. effect of steps

Active Publication Date: 2019-08-02
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because it judges the result by observing the agglutination reaction with the naked eye, the sensitivity is not high, and it cannot detect the detection objects that require high sensitivity.

Method used

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  • Exclusion chromatography detection new method of immune complex
  • Exclusion chromatography detection new method of immune complex
  • Exclusion chromatography detection new method of immune complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] see image 3 , provide a kind of exclusion chromatography detection method of hepatitis B surface antigen, the steps are as follows:

[0045] (1) Coating the hepatitis B surface antibody on the surface of the carrier particles, and labeling the paired hepatitis B surface antibody with fluorescein;

[0046] (2) Add the sample 1 to be tested, the carrier particles 2 coated with HBsAb and the fluorescein-labeled HBsAb 3 into the upper large column 20 at the same time, and incubate at 37° C. for 1 hour;

[0047] (3) Centrifuge, so that the carrier particles pass through the molecular sieve chromatography medium 4 to reach the bottom of the lower small column 22, and interpret the result by an instrument (such as a fluorescence microscope). If there is fluorescein on the surface of the carrier particle at the bottom of the lower small column 22, it is positive 5; if there is no fluorescein on the surface of the carrier particle at the bottom of the lower small column 22, it i...

Embodiment 2

[0049] see Figure 4 , provide a syphilis antibody size exclusion chromatography detection method, the steps are as follows:

[0050] (1) Coating the syphilis antigen on the surface of the carrier particles, and marking the paired syphilis antigen with quantum dots;

[0051] (2) Add the sample to be tested 1, the carrier particle 2 coated with syphilis antigen and the syphilis antigen 3 labeled with quantum dots to the upper large column 20 at the same time, and incubate at 37° C. for 1 hour;

[0052] (3) Centrifuge, so that the carrier particles pass through the molecular sieve chromatography medium 4 to reach the bottom of the lower small column 22, and interpret the result by an instrument (such as an ultraviolet imager). If the surface of the carrier particle at the bottom of the lower small column 22 has quantum dots, it is positive 5; if the surface of the carrier particle at the bottom of the lower small column 22 has no quantum dots, it is negative 6.

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Abstract

The invention discloses an exclusion chromatography detection new method of an immune complex. A double antigen or double antibody sandwich method is used to carry out detection. The method comprisesthe following steps of 1) selecting carrier particles, coating detection antigens / antibodies on a surface of the carrier particles, and marking the paired antigens / antibodies with markers; 2) simultaneously adding the carrier particles coated with the detection antigens / antibodies, the antigens / antibodies with the markers, and a sample to be tested to a reaction container; 3) incubating and centrifuging; and 4) detecting through a detector. Compared with traditional ELISA and CLIA, in the exclusion chromatography detection new method of the immune complex, a washing step is omitted, operationis simple and detection time is shortened. The antigens and antibodies belong to a same phase reaction in a microcolumn cavity, the reaction is sufficient so as to avoid a disadvantage of an insufficient ELISA heterogeneous reaction. Compared with MGIA, fluorescence and other immunolabeling verification is performed, a signal is amplified, sensitivity is greatly increased and an application rangeis wide.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a new method for detecting immune complexes by exclusion chromatography. Background technique [0002] Immunodiagnosis is a method for detecting various diseases based on the specific immune reaction between antigens and antibodies. Immune complex, also known as antigen-antibody complex, is the product of the combination of antigen and antibody, and is the detection object of immunodiagnosis. Usually, known antibodies or antigens are labeled with easy-to-display substances, and the markers are detected to reflect the presence or absence of antigen-antibody reactions, thereby indirectly measuring trace amounts of antigens or antibodies. Commonly used markers include enzymes, fluoresceins, radioactive isotopes, colloidal gold, and electron-dense substances. The specific reaction carried out by the display on the antigen or antibody label is called immunolabeling tech...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/559
CPCG01N33/559
Inventor 李勇段生宝丁少华陈晔洲王红梅魏双施谢劲松李冬李剑平刘永茂
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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