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Anti-rabies virus G protein scFv20 antibody and application thereof

A technology of rabies virus and scfv20, applied in the direction of antiviral immunoglobulin, antiviral agent, application, etc., can solve the problems of heterogeneous protein allergic reaction, antibody quality cannot be guaranteed, low yield, etc., to achieve high specificity and affinity, guarantee Variety of storage capacity, easy to penetrate the effect of tissue

Pending Publication Date: 2021-03-02
NANJING MEDICAL UNIV
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AI Technical Summary

Problems solved by technology

Since HRIG needs to be extracted from the serum of rabies vaccinators, the quality of the antibody cannot be guaranteed, the cost is high, and the output is low, so it is difficult to meet the needs of clinical medication; while ERIG has problems such as easy to cause allergic reactions to heterologous proteins, and the safety of medication cannot be guaranteed.

Method used

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  • Anti-rabies virus G protein scFv20 antibody and application thereof
  • Anti-rabies virus G protein scFv20 antibody and application thereof
  • Anti-rabies virus G protein scFv20 antibody and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0025] RVG protein gene amplification, protein expression, purification and identification:

[0026] According to the G protein gene sequence of rabies virus CVS-11 strain included in GenBank, specific primers were designed, and the target gene was amplified with cDNA as a template, with a size of 1600bp ( figure 1 ), the PCR product was purified and ligated with a plasmid vector, and the sequenced correct recombinant plasmid was transformed into Escherichia coli Top10F', induced by IPTG, and purified to obtain G protein with a size of 67 000. The results of SDS-PAGE and Western blot are shown in Figure 2-3 .

[0027] Establishment of fully human anti-rabies virus scFv phage antibody library:

[0028] Using the cDNA obtained by reverse transcription as a template, the size of the VL and VH genes was amplified by PCR to 350-400bp, and then the scFv gene was obtained by overlappingPCR with a size of 750bp ( Figure 4 ). The scFv gene and vector pComb3xss were ligated and ele...

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Abstract

The invention provides an anti-rabies virus G protein scFv20 antibody and application thereof. A light chain amino acid sequence is shown as SEQ ID NO.1, and a heavy chain amino acid sequence is shownas SEQ ID NO.2. According to the invention, a phage antibody library technology is applied, 130 cases of peripheral blood lymphocytes of healthy volunteers immunized by rabies vaccines are used to construct a fully humanized anti-rabies virus scFv phage antibody library, and purified RVG protein is used as an antigen to enrich and screen the phage antibody library to obtain a strain of fully humanized anti-RVG protein scFv with neutralizing activity, named scFv20; wherein the neutralization titer is determined to be 0.5 IU / mg.

Description

technical field [0001] The invention belongs to the technical field of antibodies, and in particular relates to an anti-rabies virus G protein scFv20 antibody and an application thereof. Background technique [0002] Rabies is a zoonotic disease caused by the rabies virus, and its fatality rate is almost 100%. Rabies kills more than 55,000 people worldwide each year, mainly in Asian and African countries. At present, the prevalence of rabies in my country is at its third peak, and it is showing a growing trend. The treatment after exposure to rabies virus mainly adopts the method of anti-rabies virus immunoglobulin (RIG) combined with rabies vaccination. There are two types of immunoglobulins currently in clinical use: equine anti-rabies virus immunoglobulin (ERIG) and human anti-rabies virus immunoglobulin (HRIG). Since HRIG needs to be extracted from the serum of rabies vaccinators, the quality of the antibody cannot be guaranteed, the cost is high, and the yield is l...

Claims

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Application Information

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IPC IPC(8): C07K16/10C12N15/13A61K39/42A61P31/14
CPCC07K16/10C07K16/005A61P31/14C07K2317/622C07K2317/76A61K2039/505
Inventor 张晓李欣妍刘方舟陆辰
Owner NANJING MEDICAL UNIV
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