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Neutralizing antibody high-sensitivity detection method and product

A detection method and a technology for detecting samples, which can be used in material inspection products, measuring devices, biological tests, etc., and can solve problems such as low neutralization ability, high mutation frequency, positive conversion rate and low titer of neutralizing antibodies

Active Publication Date: 2021-07-23
GUANGDONG FAPON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, some vaccines show signs of low positive conversion rate and titer of neutralizing antibodies after immunization
In addition, as an RNA virus, SARS-CoV-2 has a very high mutation frequency, and the neutralizing effect of convalescent serum and vaccines shows lower neutralization ability for some mutations, and the accumulation of these mutations may lead to the occurrence of immune escape

Method used

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  • Neutralizing antibody high-sensitivity detection method and product
  • Neutralizing antibody high-sensitivity detection method and product
  • Neutralizing antibody high-sensitivity detection method and product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] 1. Preparation of colloidal gold

[0093]The colloidal gold solution is prepared by the following method: chloroauric acid is formulated into a 1% solution, and then the above-mentioned solution with a final concentration of 4 / 10,000 is added to the boiled purified water, and the boiling is continued for 3 minutes, and 0.1M sodium citrate is added according to Add 580 μL per 100 mL of chloroauric acid solution, continue stirring and heating for 10 minutes, and cool to room temperature to prepare a colloidal gold solution, which is stored at 2-8°C for later use.

[0094] 2. Colloidal gold labeling

[0095] Using the SARS-CoV-2S protein RBD antibody (No. G3, available from Guangdong Feipeng Biological Co., Ltd.), which can compete with human ACE2, the steps of connecting colloidal gold are as follows: add K to the colloidal gold solution 2 CO 3 , adjust the pH to 8.0, add G3 to the adjusted pH solution to make the protein content 10μg / mL, couple for 5min, add 10% BSA to...

Embodiment 2

[0110] 1. Preparation of colloidal gold

[0111] The colloidal gold solution is prepared by the following method: chloroauric acid is formulated into a 1% solution, and then the above-mentioned solution with a final concentration of 4 / 10,000 is added to the boiled purified water, and the boiling is continued for 3 minutes, and 0.1M sodium citrate is added according to Add 580 μL per 100 mL of chloroauric acid solution, continue stirring and heating for 10 minutes, and cool to room temperature to prepare a colloidal gold solution, which is stored at 2-8°C for later use.

[0112] 2. Colloidal gold labeling

[0113] The SARS-CoV-2S protein RBD antibody G3 used, the steps of connecting colloidal gold are as follows: add K to the colloidal gold solution 2 CO 3 , adjust the pH to 8.0, add G3 to the adjusted pH solution to make the protein content 10μg / mL, couple for 5min, add 10% BSA to it to terminate the coupling reaction; discard the supernatant after centrifugation, and rediss...

Embodiment 3

[0130] 1. Preparation of colloidal gold

[0131] The colloidal gold solution is prepared by the following method: chloroauric acid is formulated into a 1% solution, and then the above-mentioned solution with a final concentration of 4 / 10,000 is added to the boiled purified water, and the boiling is continued for 3 minutes, and 0.1M sodium citrate is added according to Add 580 μL per 100 mL of chloroauric acid solution, continue stirring and heating for 10 minutes, and cool to room temperature to prepare a colloidal gold solution, which is stored at 2-8°C for later use.

[0132] 2. Colloidal gold labeling

[0133] Using human ACE2 protein Ag2, the steps of connecting colloidal gold are as follows: add K to the colloidal gold solution 2 CO 3 , adjust the pH to 8.0, add Ag2 to the adjusted pH solution to make the protein content 10 μg / mL, couple for 5 minutes, add 10% BSA to it to terminate the coupling reaction; discard the supernatant after centrifugation, and redissolve the ...

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Abstract

The invention relates to the field of antibody detection, in particular to an antibody detection method and product which have higher sensitivity when applied to neutralizing antibody detection, can further detect total antibodies and have wide market application value in current novel coronavirus neutralizing antibody and total antibody detection. The detection method provided by the invention comprises the step of contacting a sample with a ligand-containing fragment, a reagent 1 and a reagent 2. Of the reagent 1 and the reagent 2, one is connected with a marker, and the other is fixed on a solid-phase carrier.

Description

technical field [0001] The present invention relates to the field of antibody detection, in particular, to antibody detection methods and products. Background technique [0002] Receptor ligand binding is one of the pathways for signal transduction, and receptors can recognize and specifically bind ligands. Taking pathogens invading host cells as an example, ligands on the surface of pathogens can bind to receptors on host cells, opening the door to infect host cells. [0003] However, in the face of brand-new viruses, under the background of many related basic researches and incomplete mechanisms, a large number of vaccine designs are rapidly entering the clinic, and there are still many gaps in the effectiveness, protection period, quality control, and accessibility of vaccines. content needs further research. [0004] The first batch of domestic vaccines were mainly inactivated vaccines. However, the inactivated BPL used in the inactivated vaccines has a high proportion...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/569G01N33/558G01N33/543
CPCG01N33/587G01N33/54346G01N33/54306G01N33/56983G01N33/558G01N2333/165G01N2469/20
Inventor 夏良雨潘少丽覃丽芳岑静邓盛花范名俊
Owner GUANGDONG FAPON BIOTECH CO LTD
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