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Cell keratin 19 fragments chemiluminescence immune analysis determination reagent kit and preparing method thereof

A chemiluminescence immunity and cytokeratin technology, which is applied in the field of immunoassay medicine, can solve the problems of limited popularization and application, cannot be widely used in clinical diagnosis and scientific research work, etc., and achieves the effect of improving sensitivity

Inactive Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The chemiluminescence immunoassay quantitative kit in the prior art is a closed 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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  • Cell keratin 19 fragments chemiluminescence immune analysis determination reagent kit and preparing method thereof
  • Cell keratin 19 fragments chemiluminescence immune analysis determination reagent kit and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of the cytokeratin 19 fragment chemiluminescent immunoassay quantitative assay kit of the present invention

[0050] 1. Preparation of enzyme-labeled cytokeratin 19 fragment CYFRA21-1 monoclonal antibody

[0051] (1) Preparation of horseradish peroxidase-labeled cytokeratin 19 fragment CYFRA21-1 monoclonal antibody

[0052] 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 of cytokeratin 19 fragment CYFRA21-1 Stir the monoclonal antibody for 2 hours, and finally reduce it with 200mmol / L NaBH4, dialyze it with 0.02M PBS buffer, add an equal volume of glycerol, and store it below -20°C.

[0053] (2) Preparation of alkaline phosphatase-labeled cytokeratin 19 fragment CYFRA21-1 monoclonal antibody

[0054] Cytokeratin 19 fragment CYFRA21-1 monoclonal antibody was coupled with alkaline phosphatase by the glutara...

Embodiment 2～3

[0110] Examples 2-3 Preparation of the cytokeratin 19 fragment chemiluminescent immunoassay quantitative assay kit of the present invention

[0111] The quantitative assay kit of the present invention was prepared in the same manner as in Example 1 except that plastic beads and magnetic particles were used as carriers respectively.

Embodiment 4

[0112] Embodiment 4 The usage method of the kit of the present invention

[0113] Before using the kit in Example 1, it is necessary to first take out the solid-phase antibody, calibrator / test sample, and the labeled antibody solution and place them at room temperature for 15-30 minutes to allow them to reach room temperature; then adjust the incubator or water bath to 37°C ; Then prepare a suitable micro-sampler and corresponding tip and check whether the chemiluminescence instrument and auxiliary equipment such as a plate washer are working normally.

[0114] Utilize this kit to carry out the operation step of experiment according to embodiment 1 as follows:

[0115] 1) Place the strips required for the experiment (antibody-coated, 96 / 48 wells can be disassembled into 12 / 6*8*1 wells) on the plate rack;

[0116] 2) Add the sample to be tested and the calibrator of each concentration in the reaction well, add 0, 1, 3, 9, 27, 81 ng / mL of each 50 μL of the calibrator to each we...

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Abstract

The invention relates to the medical field of immunoassay, more specially, the invention provides a cytokeratin 19 fragment chemiluminescent immunoassay detection kit and a preparation method thereof. The kit of the invention comprises 1) cytokeratin 19 fragment CYFRA21-1 calibrators, 2) solid-phase vectors which are coated by cytokeratin 19 fragment CYFRA21-1 monoclonal antibodies, 3) cytokeratin 19 fragment CYFRA21-1 monoclonal antibodies which are marked by enzyme and 4) chemiluminescent substrates which are acted by the enzyme. Further, the preparation method of the kit according to the invention comprises the following steps: 1) preparing the cytokeratin 19 fragment CYFRA21-1 calibrators, 2) coating the solid-phase vectors with the cytokeratin 19 fragment CYFRA21-1 monoclonal antibodies, 3) marking the cytokeratin 19 fragment CYFRA21-1 monoclonal antibodies with the enzyme, 4) packaging the calibrators, the enzyme markers and the chemiluminescent substrates and 5) assembling finished products. The kit of the invention has the advantages of convenience, rapidness, sensitivity, stability and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a cytokeratin 19 fragment chemiluminescent immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Lung cancer is characterized by high malignancy and rapid disease progression. In recent years, its incidence has been on the rise, and it is one of the common tumors that endanger human health and cause human death. Lung cancer can be divided into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). NSCLC is the most common type of lung cancer, accounting for about 70% to 80% of lung cancer cases, and the five-year survival rate of lung cancer at each stage is only 13%. Therefore, improving the diagnosis level of lung cancer is of great significance to the timely treatment and prognosis monitoring of patients. [0003] Soluble cytokeratin 19 fragment (cytokeratin 19 fragment, CYFRA21-1) is a n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N21/76G01N33/574
Inventor 石根林金明王栩应希堂胡国茂
Owner CHEMCLIN DIAGNOSTICS CO LTD
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