Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of high-positive-charge green fluorescent protein in preparing liver cancer early diagnostic kit

A green fluorescent protein, high positive charge technology, applied in the field of medicine and cell biology, can solve the problems affecting the specificity of HCC diagnosis, missed tumor detection, etc., and achieve the effect of saving measurement time, simple steps, and reducing the amount of use

Inactive Publication Date: 2017-08-22
山东大学深圳研究院
View PDF6 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, alpha-fetoprotein (AFP) is a molecular marker of HCC widely used clinically, but in the early diagnosis of HCC, if only relying on the detection of AFP in serum, tumors with a diameter of 5 cm and smaller will be missed
In addition, AFP levels are also significantly elevated in the serum of a large number of patients with benign liver disease, which greatly affects its specificity in the diagnosis of HCC

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of high-positive-charge green fluorescent protein in preparing liver cancer early diagnostic kit
  • Application of high-positive-charge green fluorescent protein in preparing liver cancer early diagnostic kit
  • Application of high-positive-charge green fluorescent protein in preparing liver cancer early diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Detection of different concentrations of GPC3 in PBS with highly positive green fluorescent protein.

[0030] 60 μL of GPC3 antibody (purchased from Abcam, 30 ng / mL) was spread on a 384 microwell white plate and blocked with 1% BSA. After blocking, wash with PBS 3 times, then add different concentrations of GPC3 and GPC3ΔGAG (GPC3 without glycosaminoglycan side chains) dissolved in PBS, leave at room temperature for 2 hours, after incubation, wash 3 times with PBS, add 0.3 μg of highly positively charged green fluorescent protein (ScGFP), placed at room temperature in the dark for 10 min, washed 5 times with PBS, and measured for fluorescence intensity (Ex: 470nm, Em: 509nm). The results showed that as the content of GPC3 and GPC3ΔGAG increased, the fluorescence intensity also increased; at the same concentration, the ability of GPC3 to bind fluorescence was stronger than that of GPC3ΔGAG (such as figure 1 shown). This shows that the glycosaminoglycan side...

Embodiment 2

[0031] Example 2. Detection of different concentrations of GPC3 in serum with highly positive green fluorescent protein.

[0032] As mentioned above, the specific antibody of GPC3 was plated in 384 microwell white plates and blocked. Then add GPC3 and GPC3ΔGAG dissolved in appropriately diluted fetal bovine serum, and place at room temperature for 2 hours. After incubation, wash 3 times with PBS, add 0.3 μg ScGFP, place in the dark at room temperature for 10 minutes, wash 5 times with PBS, and measure the fluorescence intensity (Ex: 470nm, Em: 509nm). The results showed that as the content of GPC3 and GPC3ΔGAG increased, the fluorescence intensity also increased; at the same concentration, GPC3 had a stronger ability to bind fluorescence than GPC3ΔGAG; but it was less sensitive than in PBS (such as figure 2 shown). This shows that the high-positive green fluorescent protein can detect GPC3 in serum, which provides a simple and quick method for the early diagnosis of liver c...

Embodiment 3

[0033] Embodiment 3, the impact of salt concentration on the detection of GPC3 in serum

[0034] Add different concentrations of NaCl to fetal bovine serum containing GPC3, incubate at 30° C. for 1 hour with occasional gentle shaking, and then detect the effect of salt concentration on the detection of GPC3 in serum by the method in Example 2. The results show that compared with no NaCl, 0.5M NaCl can significantly improve the fluorescence intensity of the detection sample (such as image 3 shown).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention relates to application of high-positive-charge green fluorescent protein in preparing a liver cancer early diagnostic kit. The diagnostic kit comprises a microwell plate where a glypican-3 (GPC3) antibody is previously fixed, high-positive-charge green fluorescent protein, hyaluronic acid / chondroitin sulfate degrading enzyme, a phosphate buffer solution and silver paper required by keeping out of the sun. According to the application of the high-positive-charge green fluorescent protein in preparing the liver cancer early diagnostic kit, the characteristic that proteoglycan is composed of two parts of core protein and side-chain glycosaminoglycan is utilized, and the proteoglycan can be quickly and flexibly detected through specific recognition of the antibody and the core protein and a charge combination effect of ScGFP and a side chain.

Description

technical field [0001] The invention relates to the application of high positive charge green fluorescent protein in the preparation of a kit for early diagnosis of liver cancer, and belongs to the technical field of medicine and cell biology. Background technique [0002] Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world, about 50% of which occur in my country, and it is the second leading cause of cancer death in my country, which seriously threatens people's health and life. The treatment of HCC is very difficult. There are often no obvious symptoms in the early stage, and it has already reached the middle and late stage when the symptoms appear. Therefore, early diagnosis has become a key and new research hotspot in the treatment of HCC, and finding HCC-specific biomarkers has become a major challenge. [0003] At present, alpha-fetoprotein (AFP) is a molecular marker of HCC widely used clinically, but in the early diagnosis of HCC, if only rel...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57438
Inventor 李福川韩乃寒王文爽王涵
Owner 山东大学深圳研究院