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Golgi protein-73 mini-ELISA detection system and use method thereof

A Golgi protein and detection system technology, which is applied in the field of Golgi protein-73 small ELISA detection system, can solve the problems of limited application, cumbersome operation, and dependence on large detection instruments, and achieve the effect of low detection cost, simple and efficient operation

Inactive Publication Date: 2017-12-05
FOSHAN YETAI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the current detection of GP73 mainly uses Western-blot, immunohistochemistry and traditional enzyme-linked immunosorbent assay (ELISA) methods. The use of medical units

Method used

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  • Golgi protein-73 mini-ELISA detection system and use method thereof
  • Golgi protein-73 mini-ELISA detection system and use method thereof
  • Golgi protein-73 mini-ELISA detection system and use method thereof

Examples

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Effect test

Embodiment 1

[0033] Embodiment 1 small-sized ELISA detection system

[0034] Golgi protein-73 (GP73) small ELISA detection system based on immunomagnetic beads, including a kit, a portable fluorescence detector disclosed in ZL201220212974.2, a microplate, a magnet plate and a magnetic bead carrier, wherein the kit includes a washing buffer , sample diluent, magnetic bead washing buffer, antibody diluent, blocking buffer, chromogenic reagent, stop solution, magnetic bead activator, GP73 standard, and horseradish peroxidase (HRP)-labeled GP73-recognizing single Cloned Antibody-5B12.

[0035] The magnetic bead washing buffer is 15Mm MES buffer at pH 6.0.

[0036] The washing buffer is PBST solution containing 0.05% Tween-20.

[0037] Both the sample diluent and the antibody diluent are PBS solutions containing 1% BSA at a pH of 7.4.

[0038] The magnetic bead activator is 15Mm MES buffer solution with pH 6.0 and 1.25M EDC solution.

[0039] The magnetic bead carrier is a carboxyl-modified...

Embodiment 2

[0044] 1) Activation of the magnetic bead carrier: take carboxyl-modified superparamagnetic microspheres as the magnetic bead carrier, wash with 15MmMES buffer at pH 6.0, add 1.25M EDC solution at a volume ratio of 1:1, and keep at room temperature After 30 minutes of optical oscillation reaction, the magnetic beads were magnetically separated using a magnet plate to obtain activated magnetic bead carriers.

[0045]2) Preparation of immunomagnetic beads coated with human GP73 monoclonal antibody 5F10: add human GP73 monoclonal antibody 5F10 diluted in a PBS solution containing 1% BSA at pH 7.4 at a ratio of 50ug antibody / mg magnetic beads, and keep away at 37°C. Light shock reaction for 2 hours, after magnetic separation and cleaning with a magnet plate, add 0.05% NaN 3 , 1% BSA in PBS solution for 30min at room temperature shaking reaction to block the free aldehyde groups of unbound antibodies, and obtain the immune activity coated with human GP73 monoclonal antibody 5F10 st...

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Abstract

The present invention discloses a Golgi protein-73 mini-ELISA detection system comprising a kit, a portable fluorescence detector, a magnet plate and a magnetic bead carrier, wherein the kit comprises a sample diluent, a washing buffer, an antibody diluent, a Blocking buffer, a chromogenic reagent, a stop solution, a bead activator, a GP73 standard, Golgi protein-73 recognization monoclonal antibody 5F10 and horse radish peroxidase-labeled Golgi protein-73 recognization monoclonal antibody 5B12. The invention also discloses a using method of the detection system. The Golgi protein-73 mini-ELISA detection system does not require the use of a microplate reader, and can be used in relevant units being lack of large equipment. The system can be used to complete the entire testing process on the basis of an immunomagnetic bead serum GP73 double monoclonal antibody sandwich ELISA test kit by combination of a portable fluorescence detector, testing cost is low, and operation is simple and efficient.

Description

technical field [0001] The invention relates to an ELISA detection system, in particular to a small-sized ELISA detection system for Golgi protein-73 (GP73). At the same time, the invention also relates to a method for using the small ELISA detection system. Background technique [0002] Liver cancer is one of the most common malignant tumors in the world, especially in my country, the incidence of liver cancer accounts for about 55% of the global total, and its mortality rate ranks second in the mortality rate of malignant tumors in my country. Because the early symptoms of liver cancer are not obvious, and it has the characteristics of rapid development and early metastasis, it is difficult to diagnose early clinically and the prognosis is poor. At present, B-ultrasound and serological detection of alpha-fetoprotein (AFP) content are the main means of liver cancer screening. However, B-ultrasound is difficult to distinguish small liver cancer and cirrhotic nodules, and A...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/574G01N33/543G01N33/535G01N21/64
CPCG01N33/68G01N21/6428G01N33/535G01N33/54326G01N33/57438G01N33/57484G01N33/577G01N2021/6439
Inventor 龙飞肖静彭涛任侃陈利军李自强
Owner FOSHAN YETAI TECH
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