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Nano antibody of Zika virus, and preparation method and application thereof

A technology of Zika virus and nano-antibody, which is applied in chemical instruments and methods, anti-viral immunoglobulin, and the use of vectors to introduce foreign genetic material, etc. It can solve the problems of sensitivity and other problems, and achieve the effect of strong specificity and high affinity

Pending Publication Date: 2019-07-05
BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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AI Technical Summary

Problems solved by technology

[0003] The present invention aims to provide a Zika virus nanobody and a preparation method thereof, to solve the sensitivity problem of Zika virus enzyme-linked immunoassay detection, so as to realize the serological detection of Zika virus among blood donors

Method used

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  • Nano antibody of Zika virus, and preparation method and application thereof
  • Nano antibody of Zika virus, and preparation method and application thereof
  • Nano antibody of Zika virus, and preparation method and application thereof

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Embodiment Construction

[0017] The present invention is further described below in conjunction with specific embodiment, so that those skilled in the art can understand the present invention, but protection scope of the present invention is not limited thereto:

[0018] 1) Preparation of Zika virus particles: use the A549 cell line to culture Zika virus, then collect the supernatant, and obtain Zika virus particles by density gradient centrifugation at a concentration of 1 mg / ml;

[0019] 2) Alpaca immunization: after the first immunization, emulsify 1mg Zika antigen with CFA, and then inject subcutaneously at multiple points; secondly, boost immunization, emulsify 0.5mg Zika antigen with IFA, and then inject subcutaneously at multiple points; finally immunize with 0.5mg Zika antigen After emulsification with IFA, subcutaneous injection at multiple points, and then serological titer verification;

[0020] 3) Target fragment cloning: first isolate peripheral lymphocytes in the blood, then perform RNA ...

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Abstract

The invention relates to the technical field of biological nano materials, in particular to a nano antibody of Zika virus, and a preparation method and application thereof. An alpaca is immunized withvirus particles of Zika virus and then cloned; cloned DNA sequences are inserted in appropriate positions of a phage coat protein structural gene, and a sequence with higher affinity is selected through affinity elutriation; finally, monoclonal ELISA (enzyme-linked immunosorbent assay) identification is carried out, and antibody expression and purification can be carried out to obtain the nano antibody of Zika virus. The nano antibody of Zika virus is obtained by immunizing the alpaca and through a phage display technology, and thus the nano antibody can be highly expressed in prokaryotic oreukaryotic systems, has strong specificity and high affinity, and is more suitable for specific serological detection of Zika virus.

Description

technical field [0001] The invention relates to the technical field of biological nanomaterials, in particular to a Zika virus nanobody and its preparation method and application. Background technique [0002] The serological detection of Zika virus can use the antibody detection of Zika virus. However, the antibodies currently on the market are generally obtained by immunizing mice or rabbits (polyclonal antibodies), or monoclonal antibodies obtained by hybridoma technology. These antibodies have the characteristics of large molecular weight and weak affinity. Contents of the invention [0003] The present invention aims to provide a Zika virus nanobody and a preparation method thereof to solve the sensitivity problem of Zika virus enzyme-linked immunoassay detection, so as to realize the serological detection of Zika virus in blood donors. [0004] In order to achieve the above object, the technical scheme adopted in the present invention is: [0005] The preparation m...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/70
CPCC07K16/10C12N15/70C07K2317/569
Inventor 陈利民李世林徐敏李玉佳段晓琼杨春晖叶海艳
Owner BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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