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Tumor-associated antigen 19-9 chemical luminescence immune analytic determination reagent kit and preparation method thereof

A tumor-related antigen, chemiluminescence technology, applied in the field of immunoassay medicine, can solve the problems of radioactive contamination, complicated operation, short storage time of reagents, etc., to achieve specific diagnosis and prognosis judgment, easy operation and production, and ensure the effect of sensitivity

Inactive Publication Date: 2009-02-25
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The basic theory of these detection techniques is generally the same, but the tracers used and the signals emitted are different, so each has advantages and disadvantages
Among them, radioimmunoassay has high sensitivity, but has disadvantages such as complex operation, unstable measurement results, short storage time of reagents, and radioactive contamination.
Enzyme-linked immunoassay overcomes many limitations of radioimmunoassay, but sensitivity is not high enough
[0006] The chemiluminescence immunoassay kit in the prior art is a closed fully automatic chemiluminescence measurement system, which requires an expensive fully automatic chemiluminescence measuring instrument, thus limiting its popularization and use, and cannot be widely used in clinical diagnosis and scientific research

Method used

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  • Tumor-associated antigen 19-9 chemical luminescence immune analytic determination reagent kit and preparation method thereof
  • Tumor-associated antigen 19-9 chemical luminescence immune analytic determination reagent kit and preparation method thereof
  • Tumor-associated antigen 19-9 chemical luminescence immune analytic determination reagent kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of the CA19-9 chemiluminescent immunoassay quantitative assay kit of the present invention

[0043] 1. Preparation of enzyme-labeled CA19-9 monoclonal antibody

[0044] The preparation of horseradish peroxidase-labeled CA19-9 monoclonal antibody is as follows:

[0045]Dissolve 4.4mg HRP in 1mL distilled water, add 0.4mL sodium periodate (50mmol / L) and stir at room temperature for 20min, dialyze through 1mmol / L sodium acetate buffer solution, pH4.4, add 8mg CA19-9 monoclonal antibody, stir 2h, finally use 200mmol / L NaBH 4 For reduction, after dialysis with 0.02M PBS buffer, add an equal volume of glycerol, and store below -20°C.

[0046] 2. Preparation of CA19-9 calibrator

[0047] Prepared with pure CA19-9, the concentrations are 0, 2, 6, 20, 60, 200U / mL, 6 bottles in total.

[0048] 3. Enzyme-labeled antibody concentration selection

[0049] The working concentration of enzyme-labeled antibody was selected by square array method as 1:6000.

...

Embodiment 2～4

[0088] Embodiment 2~4 prepares CA19-9 chemiluminescent immunoassay quantitative assay kit of the present invention

[0089] Except that plastic beads, plastic tubes, and magnetic particles were used as carriers, the CA19-9 chemiluminescent immunoassay quantitative assay kit was prepared in the same manner as in Example 1.

Embodiment 5

[0090] Embodiment 5 The using method of kit of the present invention

[0091] The specific operations of the CA19-9 chemiluminescent immunoassay quantitative assay kit prepared in the above example 1 are as follows:

[0092] 1) Take out the kit from the refrigerator at 4°C, and equilibrate at room temperature for 15 minutes;

[0093] 2) Take out the coated slats and insert them into the plate frame;

[0094] 3) Add each concentration of calibrator (0, 2, 6, 20, 60, 200 U / mL) and 25 μL of the sample to be tested in the reaction wells, set a blank well for each experiment, and then add enzyme to each well except the blank well 100 μL of markers, shake and mix thoroughly with a micro shaker, and incubate at 37°C for 1 hour;

[0095] 4) Shake off the reaction solution, fill each well with diluted washing solution, wash the plate 5 times, and finally dry it on clean absorbent paper;

[0096] 5) Add 100 μL of chemiluminescent substrate solution to each well, shake and mix well wi...

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Abstract

The invention relates to the immune assay medical field, and in particular provides a tumor-associated antigen 19-9 chemiluminescence immune assay determination kit and a preparation method thereof. According to the invention, the kit comprises (1) a CA19-9 calibrator, (2) a CA19-9 monoclonal antibody envelop vector, (3) a horse radish peroxidase marker of a CA19-9 monoclonal antibody and (4) a chemiluminescence substrate. Furthermore, the preparation method of the invention comprises the following steps: (1) the calibrator is prepared with pure CA19-9; (2) the vector is enveloped by the CA19-9 monoclonal antibody; (3) the CA19-9 monoclonal antibody is marked by the horse radish peroxidase; (4) the hemiluminescence substrate is prepared; (5) the CA19-9 calibrator, the peroxidase marker and the chemiluminescence substrate are split-charged; and (6) the finished product is assembled. The kit has the advantages of convenience, quickness, sensitivity, specificity and stability, etc.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a tumor-associated antigen 19-9 chemiluminescence immunoassay assay kit and a preparation method thereof. Background technique [0002] Tumor is a kind of malignant disease that seriously threatens human life and health. WHO pointed out that early diagnosis and treatment is one of the effective means to fight tumors. [0003] There are more than one hundred kinds of tumor markers discovered so far. Since the content of tumor markers in the body is very low in most cases, it is difficult to accurately quantify them with general detection methods. Tumor-associated antigen 19-9 (CA19-9) is a mucus-type glycoprotein, which has a high value in the diagnosis of digestive system tumors such as pancreas, stomach, and colon. Therefore, how to effectively apply CA19-9 in tumor diagnosis has always been a problem that people pay attention to and hope to solve. ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574G01N21/76
Inventor 任世奇王栩林金明李振甲应希堂胡国茂
Owner 北京科美东雅生物技术有限公司
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