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Method for immunological detection by combining acridinium ester labeling technology with general magnetic particles

A technology of immunological detection and labeling technology, which is applied in the field of immunological assay kits, and can solve the problems of multiple manpower and material resources, consumption, and impact on production efficiency

Inactive Publication Date: 2012-07-11
苏州长光华医生物试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for a long time, people have used magnetic particles as solid-phase carriers, simply taking advantage of the easy separation of magnetic particles. Most applications just label different molecules directly on the magnetic particles to participate in the immune response. Each project requires different Antigens / antibodies are labeled on magnetic particles. For large-scale production, this will consume a lot of manpower and material resources, and will seriously affect production efficiency. The perfect combination of properties and easy separation of superparamagnetic particle system, (streptavidin) is coupled with superparamagnetic particles to form a universal carrier, which can be combined with any biotinylated molecule and detected by acridinium ester The high sensitivity and high stability of the system, the development of different immunological detection kits

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation method of acridinium ester chemiluminescent detection kit for quantitative detection of alpha-fetoprotein in serum with high sensitivity

[0026] The preparation method of the present embodiment comprises the following steps:

[0027] A. Preparation of general magnetic particles:

[0028] Preparation of magnetic particle washing buffer: use double distilled water and 2-(N-phenoline)ethanesulfonic acid (MES) to prepare MES buffer solution with a pH value of 4.7-5.0 and a concentration of 50mmol / ml, add Tween-20 to the final The concentration is 0.15%, the final concentration of Proclin-300 is 0.1%, and it is sterilized by 0.22 μm microporous membrane and stored at 2-8°C for later use, and the validity period is three months.

[0029] Preparation of labeling buffer: use double distilled water, boric acid and borax to prepare a boric acid buffer with a pH of 8.3-8.7 and a final concentration of 50 mmol / ml, add Tween-20 to a final concentration of 0.15%, and a ...

Embodiment 2

[0052] Preparation method of acridinium ester chemiluminescence detection kit for combined detection of HIV-1+2 antibody and P24 antigen in blood

[0053] The preparation method of the present embodiment comprises the following steps:

[0054] A. Preparation of general magnetic particles:

[0055] Preparation of magnetic particle washing buffer: same as in Example 1.

[0056] Preparation of labeling buffer: same as Example 1.

[0057] The preparation of working buffer: with embodiment 1.

[0058] The preparation method of neutravidinized universal magnetic particles is as follows: wash the magnetic particles (0.5-5um) with magnetic particle washing buffer buffer, add EDC and NHS to make the final concentration of both 50mmol, and react at room temperature for 30-60 minutes , after washing 3 times, add neutravidin to make the molar ratio of neutravidin to magnetic particles 3-5:1, react at room temperature for 3 hours, add 50mM PBS containing 0.5% BSA to block at room temper...

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Abstract

The invention relates to the field of analyzing immunological determination, in particular to a method for immunological detection by combining an acridinium ester labeling technology with general magnetic particles and an immunological determination (quantitative / qualitative) kit which is prepared by utilizing the method. According to the method disclosed by the invention, substances, such as acridinium ester or acridine sulfonamide are used for labeling and detecting an antigen or an antibody, biotin is used for labeling and capturing the antigen or the antibody, and the immunological determination of biomolecules is carried out by combining the general (chain enzyme) avidin labeled magnetic particles. The method disclosed by the invention has the advantages of high sensitivity, wide detection range and capabilities of realizing the automation easily and having a wide application prospect in the field of the immunological detection.

Description

technical field [0001] The invention relates to the field of immunological determination and analysis, in particular to a method for immunological detection using acridinium ester labeling technology combined with general-purpose magnetic particles, and an immunological determination (quantitative / qualitative) kit prepared by the method. The invention combines acridinium ester chemiluminescence technology, biotin labeling technology and general magnetic particle technology coupled with (streptase) avidin. Background technique [0002] There are many kinds of common chemiluminescent systems, the most important ones are: HRP-luminol system (or isoluminol and its derivatives), AP-spiroadamantane-1,2-dioxane and its derivatives system, acridinium ester or acridine sulfonamide system, and electrochemiluminescence system, etc. Others that have been published in the literature but have not been widely used in commercial applications include imidazole compounds, glucose oxidase, aro...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
Inventor 姚洪涛
Owner 苏州长光华医生物试剂有限公司
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