Antigen detection kit, preparation method for antigen detection kit, and antigen detection method

An antigen detection and kit technology, applied in the field of biochemistry, can solve problems such as inaccurate antigen detection results, and achieve the effects of avoiding steric hindrance effects, accurately determining antigen content, and ensuring binding rate.

Pending Publication Date: 2020-05-01
NINGBO RUI BIO TECH
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Problems solved by technology

[0005] The purpose of the embodiments of the present invention is to provide an antigen detection kit, which aims to solve the technical problem of inaccura

Method used

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  • Antigen detection kit, preparation method for antigen detection kit, and antigen detection method
  • Antigen detection kit, preparation method for antigen detection kit, and antigen detection method
  • Antigen detection kit, preparation method for antigen detection kit, and antigen detection method

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preparation example Construction

[0035] The embodiment of the present invention also provides a method for preparing an antigen detection kit, which includes the following steps: Step S102, adding biotin to the monoclonal antibody solution containing only Fab fragments for reaction.

[0036] Step S104, after adding phosphate buffer, perform ultrafiltration to remove free biotin in the solution.

[0037] Step S106, after concentration by ultrafiltration, dilute with a buffer matrix, and add monoclonal antibodies containing Fab fragments and Fc fragments after dilution to obtain R1 solution.

[0038] Step S108, adding the chemiluminescence label to the secondary antibody solution for reaction.

[0039] Step S110, after adding phosphate buffer, ultrafiltration to remove free chemiluminescent markers in the solution.

[0040] Step S112, after concentration by ultrafiltration, use buffer matrix to dilute, add streptavidin magnetic beads to obtain R2 solution.

[0041] In the embodiment of the present invention, ...

Embodiment 1

[0066] (1) Preparation of procalcitonin (PCT, PCT is used instead of procalcitonin) detection kit:

[0067] Add 1mg of biotin-NHS to 20mg of rabbit anti-human PCT monoclonal antibody, and incubate at 37°C for 30 minutes;

[0068] Use a 10mM / L phosphate buffer with a pH value of 7.0 to remove free biotin by ultrafiltration, and after ultrafiltration and concentration, use a buffer matrix to dilute to a biotin-labeled rabbit anti-human PCT monoclonal antibody concentration of 1 μg / mL. Buffer matrix includes phosphate buffer 10mM / L, bovine serum albumin 10g / L, Tween-20 5g / L, sodium chloride 8g / L and sodium azide 0.2g / L;

[0069] Add mouse anti-human PCT monoclonal antibody to the diluted buffer matrix until the concentration of mouse anti-human PCT monoclonal antibody is 1 μ g / mL to prepare R1 solution;

[0070] Add 1 mg of acridinium ester-NHS to 20 mg of goat anti-mouse IgG secondary antibody and incubate at 37°C for 30 minutes;

[0071] Use 10mM / L phosphate buffer with a pH ...

Embodiment 2

[0087] Identical to other steps of Example 1, the difference is only that (1) the steps of preparation of the procalcitonin detection kit are specifically:

[0088] Add 1 mg of biotin-NHS to 20 mg of rabbit anti-human PCT monoclonal antibody, and incubate at 37°C for 120 minutes;

[0089] Use a 10mM / L phosphate buffer with a pH value of 7.0 to remove free biotin by ultrafiltration, and after ultrafiltration and concentration, use a buffer matrix to dilute to a biotin-labeled rabbit anti-human PCT monoclonal antibody concentration of 100 μg / mL. Buffer matrix includes phosphate buffer 10mM / L, bovine serum albumin 10g / L, Tween-20 5g / L, sodium chloride 8g / L and sodium azide 0.2g / L;

[0090] Add mouse anti-human PCT monoclonal antibody to the diluted buffer matrix until the concentration of mouse anti-human PCT monoclonal antibody is 100 μ g / mL to prepare R1 solution;

[0091] Add 1 mg of acridinium ester-NHS to 20 mg of goat anti-mouse IgG secondary antibody and incubate at 37°C ...

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Abstract

The invention belongs to the technical field of biochemistry, and particularly relates to an antigen detection kit, a preparation method for the antigen detection kit, and an antigen detection method.The antigen detection kit comprises an R1 solution and an R2 solution, wherein the R1 solution comprises a monoclonal antibody containing Fab fragments and Fc fragments, and a biotin-marked monoclonal antibody only containing Fab fragments or a biotin-marked monoclonal antibody that is not homologous to the monoclonal antibody containing the Fab fragments and the Fc fragments; and the R2 solutioncomprises streptavidin magnetic beads, and a secondary antibody marked by a chemiluminescent marker. The antigen detection kit provided by the embodiment of the invention has the advantages that structures of the two monoclonal antibodies in the R1 solution are both undamaged, and magnetic microspheres are dispensed with, so that good antigen and antibody binding effects can be ensured, and the antigen content can be accurately measured with assistance of the R2 solution.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and in particular relates to an antigen detection kit, a preparation method thereof and an antigen detection method. Background technique [0002] In the field of biochemistry, it is usually necessary to use the antibody corresponding to the antigen for antigen detection. For the convenience of operation, the antibody corresponding to the antigen is first prepared into an antigen reagent detection box. During the detection process, only the antigen reagent detection box needs to be added. Antigen detection can be completed. [0003] Most of the existing antigen reagent detection kits are based on the double-antibody sandwich method. Usually, the two monoclonal antibodies are modified with chemical substances, that is, the two monoclonal antibodies are coupled with tracer substances and magnetic microspheres respectively, and then added In the process of antigen detection, an antibody (cou...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/532G01N33/543G01N33/74G01N33/68
CPCG01N33/577G01N33/532G01N33/54326G01N33/74G01N33/68G01N2333/4712G01N2333/72
Inventor 周广亮陈媛章思思徐孝锋张闻周海滨
Owner NINGBO RUI BIO TECH
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