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Kit for detecting Hepassocin (HPS)

A kit and a technology for detecting antibodies, which are applied in measuring devices, instruments, scientific instruments, etc., to achieve high sensitivity, high specificity, and broad application prospects

Active Publication Date: 2012-03-28
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, domestic and foreign studies on this gene are mostly focused on the physiological function and mechanism of action. The existing research results show that: (1) the expression of HPS is tissue specific, and it is specifically expressed in liver tissue; (2) the effect of HPS is specific, and it can Promote the proliferation of normal liver cell line L02 by activating the MAPK / ERK signaling pathway, but have no stimulating effect on the proliferation of liver cancer cell lines and other tissue cell lines; (3) the expression is up-regulated in the process of liver injury

Method used

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  • Kit for detecting Hepassocin (HPS)
  • Kit for detecting Hepassocin (HPS)
  • Kit for detecting Hepassocin (HPS)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, the composition of kit

[0041] 1. Composition

[0042] 1. Enzyme plate coated with coated antibody

[0043] The microplate is prepared according to the following method:

[0044] Dissolving the coating antibody in the coating buffer until the concentration of the coating antibody is 2 μg / ml to obtain a mixture;

[0045] Add the mixed solution into the wells of the microplate, 0.1ml per well, and place it at 37°C for 2 hours or at 2-8°C for 16-20 hours; then add the blocking solution to each well, 0.2ml per well ml, placed at 37°C for 2 hours; placed in a drying room at 25-35°C, humidity <40%, drying time 20-24 hours, to obtain an ELISA plate.

[0046] The coating antibody is HPS monoclonal antibody, product name is 1C8, purchased from Beijing Proteome Research Center, product catalog number is AB2010-002.

[0047]The composition of the coating buffer: the solvent is 0.05M PH9.6CB buffer, the solute and its mass concentration in the coating buffer are ...

Embodiment 2

[0093] Embodiment 2, the effect of kit

[0094] 1. Measuring range: 10~1000ng / ml.

[0095] Detection method: Dilute HPS antigen to 10, 20, 50, 100, 200, 500, 1000, 2000, 4000ng / ml to obtain HPS series calibrator. Add 100 μl serial calibrators to each well, shake gently to mix. Incubate at 37°C for 30 minutes. Wash the plate 5 times, add 100 μl of enzyme conjugate (ie, enzyme-labeled antibody) to each well, and incubate at 37° C. for 30 minutes. Wash the plate 5 times, add 50 μl each of chromogenic solution A and B to each well, and develop color at 37°C in the dark for 15 minutes. Add 50 μl of stop solution to each well, and use 450 / 630 nm dual-wavelength detection on a microplate reader.

[0096] The results are shown in Table 1, where the detection range of the calibrator is 10 to 1000, and the fitting curve with a correlation coefficient of 0.999 is shown in figure 2 shown.

[0097] Table 1. Experimental results of the assay range of the kit

[0098]

[0099] Fro...

Embodiment 3

[0130] Embodiment 3, for the diagnosis of liver cancer

[0131] Clinical samples were collected from the 302 Hospital. A total of 482 normal human serum samples, 236 serum samples from patients with benign liver diseases, and 171 liver cancer samples were collected. The HPS content of each serum sample was detected by HPS quantitative assay kit (enzyme-linked immunoassay).

[0132] (1) Distribution statistics of measured values ​​of normal samples

[0133] Concentration range (ng / ml)

Number of cases

＜10

7

10～30

16

30～50

61

50～70

104

70～90

114

90～110

96

110～130

55

130～150

22

＞150

7

Total

482

[0134] (2) Statistics of measured values ​​of samples from patients with benign liver disease

[0135] Concentration range (ng / ml)

Number of cases

＜50

15

50～70

12

70～90 ...

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Abstract

The invention discloses a kit for detecting Hepassocin (HPS), which comprises an anti-Hepassocin antibody 1 and an anti-Hepassocin antibody 2, wherein the antibody 1 serves as a coating antibody, and the antibody 2 serves as a detection antibody. The kit disclosed by the invention has high sensitivity and high specificity and has a wide application prospect in the field of the quantitative detection of the HPS. The kit disclosed by the invention can be used for detecting liver injury related diseases, and also has certain clinical significance on diagnosis of liver cancer related diseases, cirrhosis related diseases and the like.

Description

technical field [0001] The invention relates to a kit for detecting HPS. Background technique [0002] Hepassocin (HPS) is a new hepatocyte proliferation-promoting factor specifically expressed in liver parenchymal cells. At present, the research on this gene at home and abroad is mostly focused on the physiological function and mechanism of action. The existing research results show that: (1) the expression of HPS is tissue-specific, and it is specifically expressed in liver tissue; (2) the effect of HPS is specific, and it can Promote the proliferation of normal liver cell line L02 by activating the MAPK / ERK signaling pathway, but have no stimulating effect on the proliferation of liver cancer cell lines and other tissue cell lines; (3) the expression is up-regulated in the process of liver injury. The expression of HPS is up-regulated after liver injury and partial hepatectomy, and can promote the proliferation of liver cells, accelerate the process of damage repair and ...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 杨晓明赵京超洪海燕许望翔曹萌萌刘晓彤李冰
Owner ACADEMY OF MILITARY MEDICAL SCI
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