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SARS coronary virus related polypeptide and its uses

A SARS virus, polypeptide coupling technology, applied in the direction of antiviral immunoglobulin, peptide, depsipeptide, etc., can solve the problem of lack of pathogens

Inactive Publication Date: 2005-02-02
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently a lack of specific antibodies against pathogens

Method used

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  • SARS coronary virus related polypeptide and its uses
  • SARS coronary virus related polypeptide and its uses
  • SARS coronary virus related polypeptide and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Artificial Synthesis of Peptides

[0052] According to the amino acid sequence shown in Table 1, the peptide was synthesized on a peptide synthesizer. After the synthesis, it was verified by mass spectrometry, which proved that the synthesized peptide was consistent with the design.

[0053] Numbering:

Embodiment 2

[0055] Linkage of the antigen polypeptide to the carrier (preparation of the conjugate)

[0056] For each polypeptide synthesized in Example 1, first dissolve it with PBS (for some insoluble polypeptides, you can first dissolve it with half an amount of PBS; 0.5mg / ml, 1mg / ml) mixed with 15mg BSA (1.5ml), then add a cross-linking agent (for different cross-linking directions, choose a different cross-linking agent, add glutaraldehyde for N-terminal cross-linking, C Add EDAC at the - end, mix and react at room temperature for 5 hours, avoid light and shake constantly; after cross-linking, put it into a dialysis bag, and dialyze against PBS at 4°C overnight; measure the volume, calculate the concentration with BSA, and store at -20°C spare.

Embodiment 3

[0058] Antigen peptide immunization animals to prepare antibodies:

[0059] Take about 0.5ml of the conjugate prepared in Example 2 (the content of the conjugate is 0.1mg and 0.5mg respectively), add 1.5ml of PBS, mix with CFA (complete Freund's adjuvant), and repeatedly push and beat in the three-way tube , until it feels laborious, after 3-5 minutes, there is no separation of the two phases.

[0060] Rabbits and mice were immunized with antigen emulsified by adjuvant; each rabbit was injected with about 3ml of antigen, and mice were injected with 0.8ml / monkey. Subcutaneously inject the bipedal pads, neck, and back of the animal into multiple points (0.2ml per point).

[0061] Three weeks after the initial immunization, take the same dose of polypeptide antigen and PBS, mix with 2ml of IFA (incomplete Freund's adjuvant), and emulsify the animals for booster immunization, and the injection dose remains unchanged.

[0062] Animals were bled three weeks later and antiserum was...

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Abstract

The present invention relates to a SARS coronary virus related polypeptide and its uses. Concretely, the invention uses polypeptide from SARS virus N albumen synthesized by manpower as antigen and the antibody (single anti, multi-anti) generated after the polypeptide is used for immunity for animals, and combines enzyme linked immunity and other methods to detect the early anti-SARS antibody IgM or IgG in human blood serum sample, or directly detect the SARS pathogen in the blood serum sample.

Description

technical field [0001] The present invention relates to SARS coronavirus-related polypeptides and applications thereof. Specifically, the present invention uses artificially synthesized polypeptides derived from the nucleocapsid protein (N protein) of SARS virus, and antibodies (mAbs, polyclonal antibodies) produced after the polypeptides are immunized with animals, combined with ELISA and other methods to detect human Early anti-SARS antibody IgM or IgG in serum samples, or directly detect SARS pathogens in serum samples. Background technique [0002] The causative agent of atypical pneumonia, Severe Acute Respiratory Syndrome (SARS), has recently been identified as a coronavirus. The virus is a positive-strand RNA virus with a genome of single-stranded RNA containing Poly(A), with a total length of nearly 30kb. The surface of the virus has a rod-shaped structure of about 20nm, which is crown-shaped. The pathogen is highly contagious and contagious. After the virus under...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C07K16/10G01N33/52G01N33/569G01N33/577G01N33/68
Inventor 孙兵吴家睿裴钢李亦学石铁流杨瑞馥何有裕施木德吕伟季永镛
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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