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Superparamagnetism composite microsphere with hepatitis C NS3 monoclonal antibody and preparation process thereof

A monoclonal antibody, superparamagnetic technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high price and inability, and achieve the effects of easy magnetic separation, improved detection sensitivity, and stable reagent properties.

Inactive Publication Date: 2010-04-14
INST OF HEPATITIS BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no reagent for detecting HCAg-NS3 in China. There are already companies in foreign countries producing kits for detecting HCV core antigen, but the price is too high to be widely used in China.

Method used

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  • Superparamagnetism composite microsphere with hepatitis C NS3 monoclonal antibody and preparation process thereof
  • Superparamagnetism composite microsphere with hepatitis C NS3 monoclonal antibody and preparation process thereof

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Experimental program
Comparison scheme
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Embodiment

[0064] 1. Preparation and storage of magnetic immune microspheres coupled with hepatitis C NS3 monoclonal antibody:

[0065] Take 100μl of avidin-conjugated magnetic microspheres in a 2ml centrifuge tube, magnetically separate, discard the supernatant, and wash with 300μl of washing solution (0.02M Tris-HCl, containing 0.02% Tween 20, pH=7.4) each time, After washing at least twice, magnetically separate, discard the supernatant, add 200μl of biotinylated hepatitis C NS3 monoclonal antibody with a concentration of 1mg / ml, mix well, shake reaction at room temperature for 20-30 minutes, and shake at 180rpm. Magnetic separation, discard the supernatant, wash 3 times with the above washing solution, add 0.5-2ml stock solution (0.2M PBS, 10% BSA, PH=7.4), store at 4°C for later use.

[0066] 2. Detection of HCAg-NS3 in serum:

[0067] Add 50μl sample diluent (0.02M PBS, 0.1% Tween 20, pH=7.4) to each test tube, then add 50μl sample to be tested, add 50μl genetic engineering HCAg-NS3 solu...

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Abstract

The invention discloses a superparamagnetism composite microsphere with hepatitis C NS3 monoclonal antibody and a preparation process thereof, and relates to the diagnosis reagent technology, in particular to a reagent for external diagnosis of magnetic immunity and a process for detecting ultramicro hepatitis C NS3 antigen in blood serum. The invention synthesizes a superparamagnetism immune microsphere whose surface is coupled with a hepatitis C NS3 monoclonal antibody by utilizing an avidin-biotin system and adopting the superparamagnetism microsphere as a carrier, then reacts the superparamagnetism immune microsphere with hepatitis C NS3 antigen in detected blood serum and hepatitis C NS 3 monoclonal antibody of enzyme label under constant temperature, operates the magnetic separation, and measures after scouring. The invention is a novel immunoreaction multistage amplifying system for detecting hepatitis C NS3 antigen, has less detection steps, can be finished the detection within one hour, has simple operation, low cost, simple requirement to experiments, high sensitiveness and detection concentration of p g / ml, and is suitable for being widely used in our country.

Description

Technical field [0001] The invention relates to the technical field of diagnostic reagents and belongs to the field of magnetic immunological in vitro diagnostic reagents. It is superparamagnetic composite microspheres coupled with hepatitis C NS3 monoclonal antibody on the surface and a preparation method thereof, and double antibody sandwich ELISA using superparamagnetic composite microspheres Method or chemiluminescence method to detect hepatitis C NS3 antigen. Background technique [0002] Hepatitis C is a major epidemic infectious disease caused by hepatitis C virus (HCV) that seriously threatens human health. At present, the main detection indicators of HCV are hepatitis C antibody (anti-HCV) and hepatitis C nucleic acid (HCV RNA). [0003] After HCV infection to anti-HCV detection, the average "window period" is 70 days, and some patients can extend the "window period" to 6-9 months or longer. About 1 to 3% of patients continue to be negative for anti-HCV , Even lifelong n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/576G01N33/531G01N33/543G01N21/76
Inventor 谢立官月平时红波闫丽武志明娄金丽韩大康董培玲
Owner INST OF HEPATITIS BEIJING
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