Novel coronavirus nucleocapsid protein antibody or antigen binding fragment for in vitro diagnosis

A technology of nucleocapsid protein and coronavirus, which is applied in the direction of antiviral immunoglobulin, antibody, antiviral agent, etc., can solve the problems of low sensitivity, prone to missed detection, prone to false positive results, etc., to achieve high sensitivity and specific effect

Active Publication Date: 2022-02-01
北京健乃喜生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main manifestations are: 1) The binding affinity between the antibody and the antigen is not high enough, resulting in low detection sensitivity, which is prone to missed detection in prac

Method used

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  • Novel coronavirus nucleocapsid protein antibody or antigen binding fragment for in vitro diagnosis
  • Novel coronavirus nucleocapsid protein antibody or antigen binding fragment for in vitro diagnosis
  • Novel coronavirus nucleocapsid protein antibody or antigen binding fragment for in vitro diagnosis

Examples

Experimental program
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Example Embodiment

[0064] Example 1

[0065] Antibodies were obtained by hybridomas. Specific steps are as follows:

[0066] Using new coronavirus nucleus proteins as antigen immunization mice. Collect mice spleen and prepare spleen cells. Mouse spleen cells were combined with Myeloma Cells and screen stable hybridoma cells. Antibodies producing antigen-specific antibodies were then screened by the method of ELISA.

[0067] Sequencing the obtained high-specific antibody. One of the antibodies is numbered MAB # 9, and the heavy chain variable region sequence is shown in SEQ ID NO: 17, and the light chain variable region sequence is shown in SEQ ID NO: 18.

[0068] The encoding sequence of the antibody is then introduced into the PCMV_HC and a PCMV_LC antibody expression vector. The expression of PCMV_HC and PCMV_LC carrier was transfected into the CHO cell line to express the expression of recombinant antibody by liposome. The purified antibody was separated by Protein A affinity column, and the puri...

Example Embodiment

[0072] Example 2

[0073] Example 2 provides a test strip, such as figure 1 Indicated. figure 1 The bid number 1 is the sample pad (for the glass fiber film), 2 is the colloidal gold pad, 3 is the nitrocellulose film, 4 is the absorbent paper, 5 is the bottom plate, 6 is the detection line (also known as a T line), 7 is quality Line (also known as C). The preparation method of each part of the test strip is as follows:

[0074] 1, preparing nitrocellulose membrane

[0075] The formulation package was buffer: containing 0.05 M pH 8.5 PBS buffer for the packaged buffer, filtered with 0.22 μm membrane, and set at 4 ° C. Among them, buffer formula: NaCl 40g, KCL 1G, NA 2 HPO 4 12h 2 O 14.5g, kh 2 PO 4 1g, double steamed deionized water to 1000 ml.

[0076] Preparation of nitrocellulose membranes: The new coronavirus nuclear shell protein recombinant antibody (MAB # 4) obtained in Example 1 was diluted with a packaged buffer to adjust the machine, and the scribe is T line; with a sheep...

Example Embodiment

[0095] Example 3

[0096] Example 3 The detection performance of the new coronavirus was verified using the test strip of Example 2:

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Abstract

The present invention provides a novel coronavirus nucleocapsid protein antibody or antigen binding fragment for in vitro diagnosis. The antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises CDR sequences as shown in SEQ ID NO: 11, 12 and 13, and the light chain variable region comprises CDR sequences as shown in SEQ ID NO: 14, 15 and 16. The antibody provided by the invention is used for in-vitro detection of novel coronavirus, and has extremely high sensitivity and specificity.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a novel coronavirus nucleocapsid protein antibody or antigen-binding fragment for in vitro diagnosis. Background technique [0002] Since the outbreak of the novel coronavirus (SARS-COV-2) in December 2019, it has spread widely around the world. After it infects the human body, it will lead to the new type of coronavirus pneumonia (COVID-19) with a high probability. It is highly contagious and pathogenic. Impact. [0003] After the outbreak of the new coronavirus, in order to realize the diagnosis of the disease and block the chain of virus transmission, a variety of new coronavirus in vitro diagnostic technologies have come out. The main methods include new coronavirus nucleic acid detection, new coronavirus antibody detection and new coronavirus antigen Several types are detected. Antibody detection is the main detection method in the early stage of the epidemic, but it is a...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13A61K39/42A61P31/14G01N33/569G01N33/577G01N33/558G01N33/58
CPCC07K16/10A61P31/14G01N33/56983G01N33/577G01N33/558G01N33/587C07K2317/565C07K2317/56C07K2317/76G01N2333/165G01N2469/10
Inventor 陈亚萍尹午山王美娜张磊贾晨晨石云敬赵雷张丛
Owner 北京健乃喜生物技术有限公司
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