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Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof

An immunoassay and natriuretic peptide technology, applied in the field of immunoassay medicine, can solve the problems of expensive instruments, radioactive contamination, and need, and achieve the effects of easy operation and production, improved sensitivity, and guaranteed sensitivity.

Inactive Publication Date: 2010-09-01
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many disadvantages in the radioimmunoassay method, such as complex operation, unstable measurement results, short storage time of reagents, radioactive contamination, expensive instruments, etc.
[0010] Chemiluminescence immunoassay is an advanced and effective method, however, the current chemiluminescence immunoassay technology has problems in the efficiency and stability of NT-proBNP immunoassay products
On the other hand, the chemiluminescence immunoassay kits in the prior art are all closed automatic chemiluminescence measurement systems, which require expensive automatic chemiluminescence measuring instruments, thus limiting the promotion and use, and cannot be widely used in clinical diagnosis and treatment. scientific research

Method used

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  • Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof
  • Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof
  • Human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of the inventive plate-type N-terminal pro-B-type natriuretic peptide quantitative detection kit

[0041] 1. Preparation of anti-NT-proBNP antibody

[0042] The NT-proBNP protein expressed recombinantly in our laboratory (disclosed in the patent application with publication number CN 101230101A) was taken out from the -80°C refrigerator, dissolved and filtered, and the protein content was determined by the Lowry method, using 20mmol / L PBS with pH 5.8 It was diluted to 0.5 mg / ml. Ten female Balb / c mice aged 6 weeks and weighing about 20 g were selected. Antigen emulsification adopts double syringe mutual pushing method. For the first immunization, the recombinantly expressed NT-proBNP protein was emulsified and mixed with an equal volume of Freund's complete adjuvant, and each mouse was injected intradermally and intraperitoneally at multiple points in the amount of 100 μg NT-proBNP. The second and third immunizations were carried out on the 14th...

Embodiment 2

[0091] Example 2 Preparation of the tubular human N-terminal pro-B-type natriuretic peptide quantitative detection kit of the present invention

[0092] Divide the horseradish peroxidase-labeled NT-proBNP monoclonal antibody, and prepare the chemiluminescent substrate, coating solution and blocking solution under the same conditions, except that plastic beads and plastic tubes are used as carriers, and the rest are the same as in Example 1. The same method was used to prepare the human N-terminal pro-B-type natriuretic peptide quantitative detection kit.

Embodiment 3

[0093] Example 3 Preparation of the magnetic particle-based human N-terminal pro-B-type natriuretic peptide quantitative detection kit of the present invention

[0094] The human N-terminal pro-B-type natriuretic peptide quantitative detection kit was prepared in the same manner as in Example 1 except that magnetic particles were used as the carrier.

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Abstract

The invention provides a human N-terminal pro-B-type natriuretic peptide (NT-proBNP) immunoassay kit, mainly comprising 1) a human NT-proBNP calibration material; 2) a vector coated with human NT-proBNP polyclonal antibodies; 3) an enzyme-labeling human NT-proBNP monoclonal antibody; and 4) an enzymatic chemical luminous substrate. The invention further provides a preparation method of the kit. On the basis of the kit of the invention, the enzyme-labeling human NT-proBNP monoclonal antibody, the human NT-proBNP polyclonal antibodies coated on the vector and human NT-proBNP antigens form a 'double-antibody sandwich' structure, thus effectively utilizing a chemiluminescence technical principle and guaranteeing detection sensitivity. In addition, the kit can be applied to an open chemiluminescence measuring instrument; and the kit has low use cost and easier popularization and application.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a human N-terminal pro-B-type natriuretic peptide (Nt-proBNP) chemiluminescent immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Cardiovascular disease is an important disease that seriously threatens human life and health no matter in our country or in the world. The World Health Organization pointed out in a communiqué issued in Geneva on the 5th "World Heart Day" on September 26, 2004 that 17 million people die from heart disease and other cardiovascular diseases every year, accounting for about It is estimated that by 2020, this number will exceed 20 million, and cardiovascular disease and stroke will become the leading causes of human death and disability. Among them, heart failure (heart failure, Heart Failure, HF) is a pathological syndrome in cardiovascular diseases that seriously endanger human ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535G01N33/543
Inventor 胡川闽易维京陈安贾向阳
Owner ARMY MEDICAL UNIV
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