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SFTSV (severe fever with thrombocytopenia syndrome virus) combinable polypeptide as well as nucleic acid coding sequence and application thereof

A technology for coding sequences and sequences, applied in the field of polypeptides that can bind to SFTSV, can solve problems such as limiting efficient neutralizing antibody screening

Active Publication Date: 2020-01-21
Y CLONE MEDICAL SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

We tested the antibody titer of SFTS patient serum in the early stage and found that the antiserum against GN protein was only diluted by about 100, which suggested that the construction of antibody library with low antibody titer SFTS patient serum may limit the high efficiency of the medium. and antibody screening

Method used

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  • SFTSV (severe fever with thrombocytopenia syndrome virus) combinable polypeptide as well as nucleic acid coding sequence and application thereof
  • SFTSV (severe fever with thrombocytopenia syndrome virus) combinable polypeptide as well as nucleic acid coding sequence and application thereof
  • SFTSV (severe fever with thrombocytopenia syndrome virus) combinable polypeptide as well as nucleic acid coding sequence and application thereof

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Embodiment Construction

[0034] 1. Preparation of Immunogen

[0035] Based on the GN protein sequence and gene sequence information of HB29 SFTSV on the NCBI website, we analyzed and designed a polypeptide sGN that can effectively induce camels to produce specific antibodies against GN protein, and linked His-tag (sGN-his) or rabbit Fc (sGN-rFc) was used for subsequent purification and detection.

[0036] 2. Camel immunity and antiserum acquisition

[0037]Bactrian camels were primed with an emulsified mixture of 250 μg sGN-rFc protein and 250 μl Freund’s complete adjuvant, and boosted with sGN-rFc protein and 250 μl Freund’s incomplete adjuvant on days 14, 28, and 42. One week after the first, second and third immunizations, blood was collected to detect the antiserum titer; one week after the fourth immunization, 200ml of blood was collected for the construction of the phage antibody library.

[0038] The antiserum titer was detected by ELISA, and the detection plate was coated with GN protein at ...

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Abstract

The invention relates to an SFTSV (severe fever with thrombocytopenia syndrome virus) combinable polypeptide which comprises three complementary determining regions CDR1-3, wherein the sequence of CDR1 is or comprises sequences shown in SEQ ID NO:1 in the description; the sequence of CDR2 is or comprises sequences shown in SEQ ID NO:2 in the description; and the sequence of CDR3 is or comprises one of sequences shown in SEQ ID NO:3 in the description. The invention aims to SFTS (severe fever with thrombocytopenia syndrome) which has a high fatality rate but the lack of effective vaccines or specific antiviral drugs and implements nano antibody drug development and research and development of diagnostic kits, through preparation of GN (glycoprotein N) protein, immune bactrian camels, a platform technology of displaying nano monoclonal antibodies through a phage library, and the like, a nano antibody VHH (variable heavy chain domain) which is specifically combined with GN is screened, CDR (complementary determining region) sequences of the nano antibody are identified, and a humanized antibody SNB is established; and meanwhile, the treatment effect of the SNB in treating SFTSV infection is assessed by using a humanized mouse model in vivo. By adopting the polypeptide, a novel potential nano antibody drug is provided for clinical treatment on SFTS, and meanwhile, a corresponding detection kit is provided for diagnosis on SFTS.

Description

[0001] priority statement [0002] This application claims the priority of the applicant's PCT application with international application number PCT / CN2019 / 097350 filed on July 23, 2019, which is incorporated herein by reference. technical field [0003] The invention relates to the field of biomedicine. More particularly, it relates to a polypeptide that can bind to SFTSV, and the application of said polypeptide in the preparation of SFTSV detection agent or SFTSV therapeutic drug. Background technique [0004] Fever with thrombocytopenia syndrome (Severe fever with thrombocytopenia syndrome, SFTS) is an acute infectious disease caused by a new type of bunya virus (Severe fever with thrombocytopenia syndrome virus, SFTSV), mainly through tick bites or the blood of acute patients , mucosal contact transmission, the main clinical manifestations are fever, thrombocytopenia, leukopenia, and multi-organ dysfunction including gastrointestinal tract, liver, and kidney. Since SFT...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C12N15/13C12N15/864A61K39/12A61P31/14G01N33/569
CPCC07K16/10G01N33/56983C07K2317/565A61P31/14C07K2317/24A61K2039/505G01N2333/08
Inventor 吴喜林吴稚伟李彦磊潘逸
Owner Y CLONE MEDICAL SCI CO LTD
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