Anti-human respiratory syncytial virus n protein antibody and immunochromatography kit using the antibody

A technology of syncytial virus and immune chromatography, applied in the field of biomedicine, can solve the problems of low titer, low specificity, and low purity

Active Publication Date: 2019-10-08
HUBEI UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its preparation method is simple and low in cost, but it has defects such as low specificity, low titer, and low purity.

Method used

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  • Anti-human respiratory syncytial virus n protein antibody and immunochromatography kit using the antibody
  • Anti-human respiratory syncytial virus n protein antibody and immunochromatography kit using the antibody
  • Anti-human respiratory syncytial virus n protein antibody and immunochromatography kit using the antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1 Preparation of two kinds of rabbit anti-human respiratory syncytial virus N protein antibodies

[0072] The preparation methods of two kinds of rabbit anti-human respiratory syncytial virus N protein antibodies are as follows:

[0073]1) After structural biological analysis and related experimental research, a short peptide composed of 14 amino acids at positions 21-34 and 14 amino acids at positions 226-239 of human respiratory syncytial virus N protein (GenBank sequence number AAB59852.1) was selected As two linear epitopes for preparing rabbit anti-human respiratory syncytial virus N protein antibody respectively, the two amino acid sequences are SKYTIQRSTGDSID and FGIAQSSTRGGSRV respectively, and these two sequences are respectively named N1 and N2;

[0074] 2) After adding a cysteine ​​to the C-terminal of N1 and the N-terminal of N2 of the amino acid sequence described in step 1), respectively synthesize and purify the polypeptides with an automatic p...

Embodiment 2

[0081] Example 2 Preparation and Application of Immunochromatography Kit Based on Quantum Dot Labeling Technology

[0082] 1. Quantum dot-labeled antibody AbN1

[0083] Add 0.4nmol carboxyl water-soluble quantum dots and 800nmol carbodiimide (EDC) to the microcentrifuge tube successively, make the volume to 1ml with MES buffer (10.66g / L MES, 0.74g / L EDTA pH 7.4), keep Mix the solution with ground and react at 37°C for 5 minutes, then add 0.34 mg of the antibody AbN1 prepared in Example 1, react in the dark for 2 hours, and add single-terminal amino polyethylene glycol (PEG2000-NH2) to a final concentration of 1% (m / v), block the unreacted activated carboxyl site, and continue to react in the dark for 1h. The reacted sample was centrifuged with an ultrafiltration tube (molecular weight cut-off 100k) at 6500rpm for 5min to a volume of 200ul. The ultrafiltered sample was transferred to an ordinary EP tube and centrifuged to remove aggregates (10000rpm, 3min). Add the supernata...

Embodiment 3

[0106] Example 3 Preparation and Application of Immunochromatography Kit Based on Colloidal Gold Labeling Technology

[0107] 1. Colloidal gold-labeled antibody AbN1

[0108] a. Preparation of 30nm colloidal gold

[0109] Take a siliconized 250ml Erlenmeyer flask, add 99ml ultrapure water, add 1% (m / v) HAuCl4 solution therein and mix evenly, heat in an oil bath and stir until boiling. 2ml of 1% (m / v) trisodium citrate aqueous solution was quickly added thereto, and the solution continued to boil for 10 min (the solution turned from blue to red during this process). Stop heating, let the solution cool down to room temperature naturally, then add ultrapure water to it to make up to 100ml.

[0110] b. Colloidal gold-labeled antibody AbN1

[0111] 1) Take a siliconized 50ml Erlenmeyer flask, add 10ml of the colloidal gold solution prepared in step a, add 240ul 0.2mol / L K to the gold solution 2 CO 3 Adjust the pH to 8.5;

[0112] 2) Add the antibody AbN1 into the colloidal go...

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Abstract

The present invention relates to anti-human respiratory syncytial virus N protein antibodies and immunochromatographic kit for detection of human respiratory syncytial virus by using the same. The anti-human respiratory syncytial virus N protein antibodies separately recognize two linear epitopes consisting of No.21-34 amino acids and No.226-239 of human respiratory syncytial virus N protein; the human respiratory syncytial virus N protein has sequence number of AAB59852.1 in GenBank; amino acid sequence of sites No.21-34 and No.226-239 of the human respiratory syncytial virus N protein are respectively SKYTIQRSTGDSID and FGIAQSSTRGGSRV. The two kinds of rabbit anti-human respiratory syncytial virus N protein antibodies have the characteristics of good specificity, high purity, high titer and low preparation cost.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to an anti-human respiratory syncytial virus N protein antibody and an immunochromatographic kit using the antibody to detect human respiratory syncytial virus. Background technique [0002] Respiratory syncytial virus (RSV) is an RNA virus belonging to the Paramyxoviridae family. The disease is transmitted by air droplets and close contact. More common in newborns and infants under 6 months. The incubation period is 3 to 7 days. Infants and young children have severe symptoms, such as high fever, rhinitis, pharyngitis and laryngitis, and later manifest as bronchiolitis and pneumonia. A small number of sick children may be complicated by otitis media, pleurisy and myocarditis. After infection in adults and older children, the main manifestation is upper respiratory tract infection. [0003] According to WHO data, there are about 64 million cases of respiratory syncytial virus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10G01N33/569G01N33/558G01N33/532
Inventor 胡征董俊杨波
Owner HUBEI UNIV OF TECH
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