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Golgi apparatus protein 73 monoclonal antibody, kit and application

A technology of monoclonal antibody and Golgi apparatus, applied in anti-animal/human immunoglobulin, biological testing, biochemical equipment and methods, etc., can solve the unfavorable early diagnosis, specificity and sensitivity of clinical diagnosis of liver cirrhosis Not advanced question

Pending Publication Date: 2020-07-07
HAINAN ZHONGSEN BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Imaging examination results are reliable, but can only be found when there are obvious changes in liver tissue structure, which is not conducive to early diagnosis
Liver biopsy can establish a definitive diagnosis, but is poorly accepted by patients due to its invasive nature
However, the current laboratory serological testing methods are difficult to give a definite diagnosis of liver cirrhosis. Taking the four tests (type III procollagen, type IV collagen, laminin and hyaluronic acid) in the stage of liver fibrosis as an example, because they are affected by Hepatic inflammation is greatly affected, and its specificity and sensitivity are not high, which cannot meet the requirements of clinical diagnosis of liver cirrhosis

Method used

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  • Golgi apparatus protein 73 monoclonal antibody, kit and application
  • Golgi apparatus protein 73 monoclonal antibody, kit and application
  • Golgi apparatus protein 73 monoclonal antibody, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This example provides a monoclonal antibody stock solution of Golgi protein 73.

[0027] The antibody stock solution is obtained by injecting the preserved hybridoma cells of the present invention in Balb / c mice to prepare ascites, and then purifying Protein G.

[0028] Specifically, ascitic fluid was prepared by injecting hybridoma cells with a deposit number of CGMCC No. 19282 in Balb / c mice, and then protein G was purified to obtain GP73 monoclonal antibody stock solution-1. Ascites was prepared by injecting hybridoma cells with the deposit number CGMCC No. 19283 in Balb / c mice, and then protein G was purified to obtain GP73 monoclonal antibody stock solution-2.

Embodiment 2

[0030] This embodiment provides a kit for detecting the level of GP73 in human serum. The specification of the kit is 96 servings / box, which contains the following components:

[0031] (1) GP73 antibody-coated plate (in each kit, 96-well plate × 1)

[0032] The coated plate is prepared by the following method: coat the GP73 monoclonal antibody stock solution-1 provided in Example 1 at a concentration of 2 μg / ml on a 96-well microtiter plate, and incubate at 2-8°C for no less than 16 hours ; Shake off the coating solution in the microplate, wash it once with phosphate buffer, add blocking solution, and incubate at 2-8°C for no less than 16 hours; shake off the blocking solution in the microplate, dry it in Store at 2-8°C.

[0033] (2) GP73 calibrator (in each kit, 0.5mL×8 bottles)

[0034] The GP73 calibrator is obtained by expressing recombinant GP73 protein (referred to as GP73 antigen) in Escherichia coli by genetic engineering methods, and then adding 0.02M phosphate buf...

Embodiment 3

[0043] This example provides a method for detecting human serum GP73 levels using the kit provided in Example 2.

[0044] 1. Sample collection: Collect venous blood and let it stand for more than half an hour. After it is fully coagulated, centrifuge at 1500rpm for 5-10min, and take the upper serum.

[0045] In order to ensure accurate detection results, the present invention requires the use of fresh samples as much as possible to avoid repeated freezing and thawing; fresh samples can be stored for 7 days at 2-8°C, and the storage period at -20°C is no more than 3 months. At -80°C, no more than 2 years; samples must be transported under cryopreservation conditions and comply with relevant biosafety regulations; samples with severe hemolysis, hyperlipidemia or serious contamination cannot be used; samples cannot use sodium azide as a preservative. If the sample has been frozen, it should be equilibrated at room temperature (18-28°C) for at least 30 minutes before testing.

[...

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Abstract

The invention relates to a Golgi apparatus protein 73 monoclonal antibody, a kit containing the antibody and application of the Golgi apparatus protein 73 monoclonal antibody in detection of Golgi apparatus protein 73. The invention also relates to a method for detecting the level of Golgi apparatus protein 73 in serum. The kit containing the GP73 monoclonal antibody is used for detecting GP73 inhuman serum, and has high sensitivity and strong specificity. Moreover, the kit and the method provided by the invention can be clinically used for auxiliary diagnosis of liver cirrhosis.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a monoclonal antibody of Golgi protein 73, a kit containing the antibody and an application in detecting Golgi protein 73. Background technique [0002] Liver cirrhosis is the advanced stage of the development of various chronic liver diseases, and it is diffuse liver damage caused by long-term or repeated effects of one or more etiologies. In my country, most of them are post-hepatitis cirrhosis, and a small part are alcoholic cirrhosis and schistosome cirrhosis. Histopathologically, extensive liver cell necrosis, nodular regeneration of residual liver cells, connective tissue hyperplasia, and fibrous septum formation lead to structural destruction of hepatic lobules and formation of pseudolobules. The liver gradually deforms and hardens and develops into cirrhosis. In the early stage, there are no obvious symptoms due to the strong compensatory function of the liver. In the...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18G01N33/68G01N33/58G01N33/577
CPCC07K16/18G01N33/6893G01N33/581G01N33/577G01N2333/47G01N2800/085
Inventor 王卫东
Owner HAINAN ZHONGSEN BIOTECH CO LTD
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