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

Method for coating mpo with solid support, product and application thereof

A solid support and coating technology, which is applied in the field of immunoassay, can solve the problems of poor reagent performance, poor MPO antibody detection effect, and low solid support encapsulation effect, achieving improved stability, high efficiency, and high precision. Effect

Active Publication Date: 2022-04-08
ZHUHAI LIVZON DIAGNOSTICS
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the current MPO antibody detection effect is poor, and the sensitivity and accuracy need to be improved. The main reason is that the performance of the detection reagent is poor, and the effect of solid support coated with MPO is low.

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
  • Method for coating mpo with solid support, product and application thereof
  • Method for coating mpo with solid support, product and application thereof
  • Method for coating mpo with solid support, product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1. Carboxylated magnetic particles coated with MPO antigen

[0055] Coating method

[0056] (1) Take 50,000 carboxylated magnetic particles (1,000 parts), magnetically separate to remove the supernatant, wash twice with 100mM MES buffer (pH 6.0), and resuspend in 152 μL MES buffer (pH 6.0);

[0057] (2) Add 28 μL of freshly prepared NHS (50 mg / mL), vortex and mix, then add 20 μL of freshly prepared EDC (50 mg / mL), shake at room temperature for 50 min (1000 rpm / min);

[0058] (3) Wash twice with 100mM borate buffer solution (pH 8.0-9.0), then resuspend with 200μL 100mM borate buffer solution (pH 8.0-9.0);

[0059] (4) Add the recombinantly expressed MPO antigen in an amount of 20ng / person, shake at room temperature for 2.5h (1500rpm / min);

[0060] (5) Using a phosphate buffer solution (pH 7.4) containing 0.5% tween-20;

[0061] (6) Wash twice with blocking solution (containing 1% BSA bovine serum albumin), add 500 μL of blocking solution, and shake at room tem...

Embodiment 2

[0063] Embodiment 2 performance test

[0064] testing method

[0065] (1) Take 50 μL of magnetic particles (including 50 magnetic particles) in a 96-well plate made of transparent material, add 50 μL of serum samples pre-diluted 50 times with PBST (containing 1% BSA bovine serum albumin), and shake at 37°C 15 minutes;

[0066] (2) Use a magnetic plate to absorb magnetic particles, remove the reaction solution, and wash 2 times with PBST;

[0067] (3) Add 50 μL of phycoerythrin-labeled anti-human IgG antibody and shake at 37°C for 15 minutes;

[0068] (4) Use a magnetic plate to absorb magnetic particles, remove the reaction solution, and wash twice with PBST;

[0069] (5) Use PBST to suspend and use an interpreter to read the fluorescence signal value.

[0070] Test Results

[0071] (1) Reactivity investigation

[0072]

[0073] From 34 samples with different anti-MPO antibody titers, it can be seen that the background of negative samples is low, the weak positive sam...

Embodiment 3

[0085] Embodiment 3, coating buffer screening

[0086] According to the coating method of Example 1, wherein, in step (3), respectively adopt 2-morpholineethanesulfonic acid (MES, pH 5.0), 4-hydroxyethylpiperazineethanesulfonic acid (HEPES, pH 8.0), phosphoric acid Coating with salt (PBS, pH 7.4), borate (pH 8.0), carbonate (CB, pH 9.6), acetate (pH 5.1) or citrate (pH 5.0) as coating buffer , and examine its reactivity, the results are shown below.

[0087]

[0088] The test results showed that the magnetic particles coated in 100mM borate buffer (pH 8.0) had the highest reactivity to weak positive, positive and strong positive samples (relative to negative samples, the fluorescence signal value was higher), and had the most Good discrimination, while the background is clean. Therefore, this coating buffer works best.

[0089] Conclusion: with 2-morpholineethanesulfonic acid (pH5.0), 4-hydroxyethylpiperazineethanesulfonic acid (pH8.0), phosphate (pH7.4), carbonate (pH9....

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

No PUM Login to View More

Abstract

The invention relates to the technical field of immunoassay, in particular, it provides a method for coating MPO with a solid support as well as its product and application. The invention provides a method for coating MPO on a solid support. In the environment of borate buffer solution, the MPO is coated on the solid support to obtain a coated product. The inventor conducted research on the MPO antigen, provided the coating method of the present invention, and found that different coating environments have different coating effects. Most of the buffers used in the prior art are mainly suitable for antibody coating. The coating efficiency for MPO is not high. When the borate buffer is used as the coating environment, the solid support has high efficiency of coating MPO, and the obtained coating product has good reaction performance, high precision and significantly improved stability.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to a method for coating MPO with a solid support and its product and application. Background technique [0002] Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a type of autoimmune vasculitis involving multiple organs such as kidney and lung. The clinical manifestations of this type of disease are complex and diverse but non-specific, and it is easy to miss and misdiagnose. The diagnosis of about 1 / 3 of the patients is delayed for more than 6 months. At the same time, the disease progresses rapidly and is dangerous. The 1-year mortality rate of untreated patients is as high as 80% %; after treatment, the 5-year survival rate of patients is about 75%. Therefore, the accurate diagnosis of AAV is extremely important clinically. As a serological marker of AAV, ANCA plays an irreplaceable role in the diagnosis of AAV, and has been included in the diagnosis...

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 Patents(China)
IPC IPC(8): C07K16/40C07K17/08C07K17/14G01N33/531G01N33/543G01N33/573
CPCC07K16/40C07K17/14C07K17/08G01N33/54306G01N33/531G01N33/573
Inventor 张鹏飞陈杰王国磊
Owner ZHUHAI LIVZON DIAGNOSTICS