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Method and kit for detecting myeloperoxidase (MPO)

A myeloperoxidase and kit technology is applied in the field of immunoassay methods and kits for detecting myeloperoxidase isotypes or subtypes, and can solve problems that do not involve the detection of MPO isotypes or subtypes.

Inactive Publication Date: 2013-02-06
杭州华得森生物技术有限公司
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Problems solved by technology

However, none of these methods involve the detection of MPO isotypes or subtypes

Method used

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  • Method and kit for detecting myeloperoxidase (MPO)

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Embodiment Construction

[0091] The present invention is further described below in conjunction with the examples.

[0092] In this example, the method of the present invention is implemented by enzyme-linked immunosorbent technology, specifically, horseradish peroxidase (HRP) is selected as the marker for labeling the antibody, and HRP substrate TMB is used as the chromogen. The kit includes solid phase plate, enzyme-labeled MPO isotype-labeled antibody, MPO calibrator, TMB chromogenic reagent, stop solution, and washing solution.

[0093] 1. Preparation of MPO subtype monoclonal antibody

[0094] Purified MPO subtype proteins were used to immunize pure-line mice, and when the mouse serum produced corresponding antibodies, the mouse splenocytes and SP2 / 0 cells could be fused. Hybridoma cells were selected by HAT and detected by enzyme-linked immunosorbent assay (ELISA). The hybridoma cells that are positive for the detection of antibodies should be cloned with limited dilution, and the positive cel...

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Abstract

The invention provides a method for detecting either homotype or subtype MPO or a composition of MPO, in particular to an immune detection method with high specificity and flexibility through one but not all of either homotype or subtype MPO or the composition of MPO. The method is used for carrying out auxiliary diagnoses on whether the patients are suffered from an anaphylactic disease, an inflammation disease, a cardiovascular disease or an autoimmune disease or not, or analyzing the risk that the prognostic patients are developed to the anaphylactic disease, the inflammation disease, the cardiovascular disease or the autoimmune disease. The invention further provides a kit for implementing the method.

Description

technical field [0001] The invention relates to a detection method, in particular to an immunoassay method and kit for detecting the isotype or subtype of myeloperoxidase. technical background [0002] Myeloperoxidase (myeloperoxidase, MPO) is an important peroxidase produced by neutrophil azurophilic granules, mainly present in neutrophils and monocytes. Mature MPO is composed of two subunits, each subunit is composed of a 59-64kDa heavy chain and a 14kDa light chain, and each subunit contains a covalently bound heme, which has MPO-specific Green spectrum characteristic. MPO can catalyze the reaction of hydrogen peroxide and chlorine to produce hypochlorous acid, which has a strong antimicrobial effect and plays a role in the production and regulation of inflammatory responses in the body. However, oxidative stress and oxidative tissue damage can result when MPO catalyzes the generation of excess oxidants. [0003] The activity of myeloperoxidase is closely related to th...

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Application Information

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IPC IPC(8): G01N33/573
Inventor 张开山苏广宇高阳
Owner 杭州华得森生物技术有限公司
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