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Magnetic particle chemiluminescence detection kit and detection method thereof

A chemiluminescence detection and detection method technology, applied in the field of medical detection, can solve the problems of reduced detection accuracy, reduced non-specific adsorption of acridine esters, reduced non-specific adsorption of acridine esters, etc., to achieve the effect of improving sensitivity

Active Publication Date: 2022-04-05
NANJING VAZYME MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is a difficulty in the application of acridinium esters in chemiluminescent immunoassays. The difficulty is that acridinium esters have certain non-specific adsorption. The bound free acridinium ester, the free acridinium ester will be adsorbed on the magnetic beads or the inner wall of the reaction, which will increase the background and reduce the detection accuracy
[0006] At present, there is no method that can reduce the non-specific adsorption of acridinium esters, and there is an urgent need to develop a method for reducing the non-specific adsorption of acridinium esters.

Method used

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  • Magnetic particle chemiluminescence detection kit and detection method thereof
  • Magnetic particle chemiluminescence detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Obtaining acridinium ester antibody

Embodiment 11

[0047] Example 1.1: Preparation of acridinium ester artificial antigen

[0048] (1) Configure 0.01mol / L PBS solution: Take a clean beaker, add 500mL of pure water to it, then add 8.78g of NaCl, 5.8g of Na 2 HPO 4 12H 2 O and 0.59g of NaHPO 4 2H 2O, adjust the pH to 7.4 with an appropriate amount of hydrochloric acid, and then adjust the volume to 1000mL.

[0049] (2) Take 200 μL of the PBS solution in step (1) into a test tube, then sequentially add 200 μg of carrier protein keyhole limpet hemocyanin (KLH) and 100 μM acridinium ester to the test tube, mix well, react at room temperature for 1 hour, and pass through 50KD ultrasonography. The filter tube was purified 5 times, and the purified product was collected into a 1.5 mL EP tube. The acridinium ester-carrier protein keyhole limpet hemocyanin coupling was completed and stored at 4°C until use.

Embodiment 12

[0050] Example 1.2: Immunization of mice with the prepared acridinium ester-carrier protein keyhole limpet hemocyanin conjugate

[0051] (1) The prepared acridinium ester-carrier protein keyhole limpet hemocyanin conjugate was mixed with Freund's complete adjuvant in equal amounts, and vortexed for 30 minutes to emulsify completely. By subcutaneous multi-point injection, the emulsion was injected into healthy BALB / c mice at a dose of 80 μg / mouse to complete the primary immunization.

[0052] (2) One month later, the acridinium ester-carrier protein keyhole limpet hemocyanin conjugate was mixed with Freund's incomplete adjuvant in equal amounts, vortexed for 30 minutes to make it completely emulsified, and injected subcutaneously at multiple points. The emulsion was injected into healthy BALB / c mice at a dose of 50 μg / mouse. That is, the second immunization is completed.

[0053] (3) The third booster immunization was carried out in the third week after the second immunizatio...

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Abstract

The invention relates to a magnetic particle chemiluminescence detection kit and a detection method thereof, the kit comprises an acridinium ester labeled antibody, a capture antibody, magnetic beads and an acridinium ester antibody, the kit and the detection method can reduce non-specific adsorption of acridinium ester, and the kit and the detection method belong to the field of medical detection. By adopting the acridinium ester antibody, non-specific adsorption of acridinium ester on the inner wall of a magnetic bead or a container is reduced, the non-specific adsorption signal value of the acridinium ester is effectively reduced by 60% or above, the chemiluminescence reaction sensitivity is improved, and practicability is achieved.

Description

technical field [0001] The invention belongs to the field of medical detection, and mainly relates to a magnetic particle chemiluminescence detection kit and a detection method thereof. Background technique [0002] According to the different luminescent substances, chemiluminescence immunoassay (CIA) can be divided into three types: direct chemiluminescence immunoassay, chemiluminescence enzyme immunoassay, and electrochemiluminescence. Among them, the direct chemiluminescent immunoassay process does not require enzyme catalysis, and directly participates in the luminescent reaction. [0003] The chemical structure of the chemiluminescent agent used in the direct luminescent immunoassay has a luminescent group, so the process does not require the catalysis of enzymes, and the luminescent reaction is directly carried out. Among them, the chemiluminescent agent directly labels the detection antibody, reacts with the sample to be tested and the magnetic bead-coated capture an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/74G01N33/577G01N21/76C07K16/44
Inventor 唐波张松峰时亚斌冯速卢佩王子鸣
Owner NANJING VAZYME MEDICAL TECH CO LTD