Kit for preparing CA125 surface Tn antigen by magnetic particle chemiluminescence immunoassay and preparation method thereof

Abstract

The invention relates to the technical field of immunoassay, in particular to a kit for preparing a CA125 surface Tn antigen by magnetic particle chemiluminescence immunoassay and a preparation methodthereof. The kit comprises a magnetic separation reagent coupled with a Fab fragment of a CA125 antibody, a biotin-lectin reagent, and a streptavidin-biotin-alkaline phosphatase coupling reagent, wherein the biotin-lectin reagent containing sialidase is capable of identifying the Tn antigen. The magnetic separation reagent is specifically bound to the CA125 antigen; the biotin-lectin specificallyrecognizes the Tn antigen of the CA125 antigen surface; the streptavidin-biotin-alkaline phosphatase coupling reagent is specifically bound to the biotin-lectin reagent to realize amplification of the detection signal; the sialidase hydrolyzes a glucosidic bond connected with the sialic acid on the Tn antigen surface and the sialic acid is released to expose the Tn antigen. Therefore, the detection accuracy, sensitivity and linear range are improved. The kit has advantages of high specificity and high sensitivity and is capable of realizing fully automated testing.

Description

technical field [0001] The invention relates to the technical field of immunodetection, in particular to a kit for magnetic particle chemiluminescence immunodetection of Tn antigen on the surface of CA125 and a preparation method thereof. Background technique [0002] Ovarian cancer ranks third in the incidence of gynecological malignancies, second only to cervical cancer and uterine body cancer, but its case fatality rate ranks first. In recent decades, although a large number of scientific researchers have devoted themselves to the research of ovarian cancer, the survival rate of this type of patients has not been significantly improved. Cancer antigen 125 (CA125) was first discovered and named by Bast et al. in 1981. It is an epitope recognized by the monoclonal antibody OC125. It was first quantified by Klug et al. in 1984 using radioimmunoassay techniques. Detect and set the normal range of CA125 <35U / ml. On the one hand, CA125 is a commonly used serum tumor marker...

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Problems solved by technology

[0007] 2) This patent oxidizes and derivatizes the coated plate, intending to solve the non-specific adsorption phenomenon of lectin VVA and antibody. Although the sugar chain of the Fc segment is blocked, it is unavoidable regardless of the oxidant or the derivatization reagent. The impact on the Fab segment, thereby affecting the activity of the antibody;

[0008] 3) This patent achieves signal amplification through the specific combination of biotin-lectin VVA and streptavidin-horseradish peroxidase. This method has uncertainty and cannot guarantee that each streptavidin Molecule binds only 1 biotin molecule, thereby impairing the final test result value;

[0009] 4) The operation steps of this patent are complicated, and the reaction time is long. The test process is eluted 12 times, and it takes nearly 4 hours to complete from the beginning of the test to the end of the test, and it is difficult to realize integration and full automation;

[0010] 5) This patent uses the OD value tested by the microplate reader as the final data for ROC curve analysis, and does not involve calibrator fitting. This processing method has certain limitations, and the final result fluctuates due to the influence of different manufacturers or models of microplate readers larger;

[0011] 6) The patent does not treat the sialic acid on the surface of the CA125 antigen, which can cover the recognition site of the lectin VVA, thereby affecting the detection sensitivity and linear range

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