Fetus alpha globulin detection particles, preparation thereof and application thereof

A globulin, fetal technology, used in biological testing, measuring devices, material inspection products, etc.

Active Publication Date: 2010-07-07
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the field of clinical detection, there is no light-induced chemiluminescence detection reagent available, especially for tumor markers and hepatitis detection items

Method used

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  • Fetus alpha globulin detection particles, preparation thereof and application thereof
  • Fetus alpha globulin detection particles, preparation thereof and application thereof
  • Fetus alpha globulin detection particles, preparation thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086]Example 1 Preparation of Luminescent Particles Coated with Anti-AFP Antibody

[0087] The improved preparation method of anti-AFP antibody-coated luminescent particles is as follows:

[0088] 1) Suspension treatment of luminescent particles: absorb a certain amount of carboxyl luminescent particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically break until the particles are resuspended, add MES buffer to adjust the luminescent particles Concentration to 100mg / ml.

[0089] 2) Antibody treatment: AFP antibody was dialyzed in 0.05M MES buffer solution with pH 6.0 (hereinafter referred to as MES buffer solution). After the dialysis was completed, the concentration was measured and adjusted to 8 mg / ml.

[0090] 3) Mix MES buffer, 100 mg / ml luminescent particle suspension (MES buffer) and 8 mg / ml anti-AFP antibody solution (MES buffer) at a volume ratio of 1:1:1, and mix quickly , to obtain ...

Embodiment 2

[0140] Example 2 Preparation of biotin-labeled antibody

[0141] Preparation:

[0142] 1) Antibody treatment: dialyze anti-AFP antibody in 0.1M NaHCO 3 solution, the antibody concentration was measured and adjusted to 1 mg / ml.

[0143] 2) Prepare 16.17mg / ml Biotin solution with DMSO.

[0144] 3) Labeling: take the processed 1 mg / ml anti-AFP antibody labeled antibody and the prepared Biotin solution, mix them in proportion, and mix them uniformly quickly. Stand at 4°C for 12-16 hours.

[0145] 4) Dialysis: Dialyze the reacted biotin-labeled antibody against biotin-labeled dialysis buffer (pH 8.0).

[0146] 5) Aspirate the dialyzed biotinylated antibody and transfer it to a clean centrifuge tube, and take a sample to determine the antibody concentration. Adjust the concentration of qualified biotin-labeled antibody to 0.5mg / ml.

[0147] React antibodies with Biotin in different ratios and detect:

[0148]

[0149] The higher the marker ratio, the greater the deviation ...

Embodiment 3

[0150] Example 3 Preparation of Photosensitive Microparticles Coated with Avidin

[0151]Photosensitive particles: use photosensitive particles with a particle size of 220±40nm (PentaTek, USA)

[0152] Preparation:

[0153] a. Treatment of photosensitive particle suspension: absorb a certain amount of photosensitive particles and centrifuge in a high-speed refrigerated centrifuge, discard the supernatant, add a certain amount of MES buffer, ultrasonically sonicate on the ultrasonic cell disruptor until the particles are resuspended, and then add MES buffer Adjust the photosensitive particle concentration to 100mg / ml.

[0154] b. Preparation of avidin solution: weigh a certain amount of Avidin, add MES buffer to dissolve to 8mg / ml.

[0155] c. Mixing: Mix the treated photosensitive microparticle suspension, 8 mg / ml Avidin and MES buffer at a volume ratio of 2:5:1, and mix quickly to obtain a reaction solution.

[0156] d. Reaction: Prepare 25mg / ml NaBH in MES buffer 3 CN so...

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Abstract

The invention relates to diagnosis reagent for fetus alpha globulin and discloses fetus alpha globulin detection particles, and in particular anti-AFP antibody-coated light-emitting particles. The invention also discloses the preparation and application of the fetus alpha globulin detection particles. In addition, the invention further discloses an in vitro detction kit for measuring fetus alpha globulin in a sample and a using method of the kit at the same time. The kit can be used in for the quantitative measurement of fetus alpha globulin in human blood serum samples and can be used in cancer auxiliary diagnosis and treatment effect monitoring of cancer patients in combination with other blood serum and clinic information.

Description

technical field [0001] The invention relates to a diagnostic reagent for cancer, in particular to fetal alpha globulin (AFP) detection particles based on the principle of light-induced chemiluminescence, its preparation and application. Background technique [0002] Immunological detection is a means of detection based on the specific reaction of antigens and antibodies. Because it can use isotopes, enzymes, chemiluminescent substances, etc. to amplify and display the detected signals, it is often used to detect proteins, hormones, etc. trace bioactive substances. [0003] Immunological testing in my country has basically experienced radioimmunoassay (emerged in the 1970s, and is still widely used in hospitals above the county level); enzyme-linked immunoassay (emerged in the 1980s, commonly used in various clinical institutions); Biomarker and immunoassay technology represented by luminescence (began to be popularized and used in the 1990s, and the product entered the growt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/52G01N21/76G01N21/64
Inventor 赵卫国王海蛟陈晨辉
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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