CN108178796A - Fever is with thrombocytopenic syndromes viral glycoprotein immue quantitative detection reagent box - Google Patents

Fever is with thrombocytopenic syndromes viral glycoprotein immue quantitative detection reagent box Download PDF

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CN108178796A
CN108178796A CN201711459224.9A CN201711459224A CN108178796A CN 108178796 A CN108178796 A CN 108178796A CN 201711459224 A CN201711459224 A CN 201711459224A CN 108178796 A CN108178796 A CN 108178796A
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monoclonal antibody
antibody
detection
fever
artificial sequence
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CN108178796B (en
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朱凤才
曾晓燕
焦永军
史凤娟
刘静娴
史智扬
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JIANGSU DISEASE PREVENTION CONTROL CENTRAL
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    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983Viruses
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/005Assays involving biological materials from specific organisms or of a specific nature from viruses
    • G01N2333/08RNA viruses
    • G01N2333/175Bunyaviridae, e.g. California encephalitis virus, Rift valley fever virus, Hantaan virus

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Abstract

Kit of the detection fever with thrombocytopenic syndromes viral glycoprotein is quantified the invention discloses a kind of, the kit includes two monoclonal antibodies with thrombocytopenic syndromes viral glycoprotein Gc for fever, and purpose of the detection fever with thrombocytopenic syndromes viral glycoprotein is realized by double antibody sandwich ELISA.The detection kit detectable concentration range of the present invention is big, and high sensitivity, high specificity, testing cost is low, easy to use, suitable for being promoted on production of vaccine field or clinical diagnosis.

Description

Fever is with thrombocytopenic syndromes viral glycoprotein immue quantitative detection reagent box
Technical field
The invention belongs to biomedical sectors, are related to a kind of fever and are quantitatively examined with thrombocytopenic syndromes viral glycoprotein Test agent box.
Background technology
Fever is with thrombocytopenic syndromes virus (severe fever with thrombocytopenia Syndrome virus, SFTSV) it is in recent years in the popular novel hemorrhagic fever viruse of one kind in China Middle Eastern.The virus People and animal are infected by the mode that tick worm (haemaphysalis longicornis, H.longicornis) bites, also can people be caused by close contact Border is propagated, and huge life, property loss are caused to area is suffered from.China has 17 provinces (city, autonomous region) and confirmed at present Also someone infects the report of the virus to the prevalence of the virus, Chinese Japan of neighbouring country, South Korea etc..
SFTSV belongs to bunyaviridae, Phlebovirus on taxology, genome by big (L), in (M), small (S) three sub-thread strand RNA segment compositions, other viruses are similar to bunyaviridae, 3 ' end of viral genome and 5 ' ends Terminal sequence is complementary.The RNA polymerase that L fragment codings RNA is relied on;M fragment coding glycoprotein precursors, through in host cell after translation The modification of protease forms two glycoprotein of Gn and Gc;S segment category ambisense RNAs are separately encoded nucleocapsid protein NP and non-structural Albumen NSs.
Prevention for disease of natural focus caused by the virus, in addition to implementing tick eradication, strengthening the strategies such as Case monitoring Outside, it is means the most basic to carry out effective vaccine inoculation to the Susceptible population for living in epidemic-stricken area.SFTSV inactivated vaccines are current Just in R&D process, and to the real-time dynamic of glycoprotein (Gn and Gc) on active ingredient-virion envelope in the vaccine, Quantitative Monitoring is that an important technical is crucial in the vaccine R&D process.
Patent of the present invention describes a kind of sugared come Gc in quantitative detection Virus culture supernatant based on double-antibody sandwich elisa The content of albumen.Since the mole ratio of two kinds of glycoprotein of the Gn on virus envelope and Gc is 1:1, and the molecular weight of the two is also big Body is suitable, therefore the content by that can obtain entire glycoprotein to the detection of Gc glycoprotein.
Invention content
One of the objects of the present invention is to provide a kind of fevers quantitatively to detect with thrombocytopenic syndromes viral glycoprotein Kit.
The second object of the present invention is that provide above-mentioned fever quantitatively detects with thrombocytopenic syndromes viral glycoprotein Coated antibody and detection antibody in kit.
The third object of the present invention is that providing a kind of detected using mentioned reagent box is generated heat with thrombocytopenic syndromes The method of glycoprotein in virus.
To achieve these goals, present invention employs following technical solutions:
The present invention provides a kind of monoclonal antibody for fever companion's thrombocytopenic syndromes viral glycoprotein Gc, institutes Monoclonal antibody is stated to include:
Light chain variable region has SEQ ID NO:Amino acid sequence shown in 2 or the ammonia with it at least 80% homology Base acid sequence;
Heavy chain variable region has SEQ ID NO:Amino acid sequence shown in 1 or the ammonia with it at least 80% homology Base acid sequence;The monoclonal antibody is named as 1C5-F5.
Preferably, the light chain variable region of 1C5-F5 antibody has SEQ ID NO:Amino acid sequence shown in 2, weight chain variable Area has SEQ ID NO:Amino acid sequence shown in 1.
The present invention provides a kind of monoclonal antibody for fever companion's thrombocytopenic syndromes viral glycoprotein Gc, institutes Monoclonal antibody is stated to include:
Light chain variable region has SEQ ID NO:Amino acid sequence shown in 4 or the ammonia with it at least 80% homology Base acid sequence;
Heavy chain variable region has SEQ ID NO:Amino acid sequence shown in 3 or the ammonia with it at least 80% homology Base acid sequence;The monoclonal antibody is named as 1H6-B10.
Preferably, the light chain variable region of 1H6-B10 antibody has SEQ ID NO:Amino acid sequence shown in 4, heavy chain can Becoming area has SEQ ID NO:Amino acid sequence shown in 3.
The antibody of conservative sequence variant including preferred antibody amino acid sequence is also included within the scope of the present invention. Conserved amino acid sequence variant includes not significantly changing the modification of the amino acid sequence of the monoclonal antibody binding property of the present invention, The variant such as substituted derived from Similar amino acids well known in the art, the missing of amino acid, increase caused by variant.
The monoclonal antibody of the present invention further includes people source and non-human source antibodies and with identical with said monoclonal antibody Function or all of transformation and optimization antibody.
The monoclonal antibody of disclosure of the invention can include one or more glycosylation positions in heavy chain and light chain variable region Point, as known in the art, one or more glycosylation sites present in variable region can cause the antibody of enhancing to be exempted from Epidemic focus changes the pharmacokinetics of antibody due to changing antigen binding.
The monoclonal antibody of the present invention can be designed as including 1 of modification, typically change antibody in Fc regions Or multiple functional characteristics, such as serum half-life, the cytotoxicity that complement combines, Fc receptors combine, and/or antigen relies on.In addition, The present invention antibody can be modified by sulphation and (e.g., one or more chemical groups can be connected to antibody) or be modified with Change its glycosylation, so as to change the one or more functions characteristic of antibody again.
Another modification that the monoclonal antibody of the present invention can be designed is Pegylation.Antibody can be by polyethylene glycol Change, so as to for example, increasing biology (such as serum) half-life period of antibody.In order to make antibody Pegylation, the antibody or its segment It is anti-with PEG usually under conditions of being suitable for one or more polyethylene glycol (PEG) group and being connected to the antibody or antibody fragment Should, such as the active ester or aldehyde derivatives of polyethylene glycol.Preferably, which is by PEG molecules (or the class with activity As reactive water-soluble polymer) carry out acylation reaction or alkylated reaction and realize.
The present invention also provides the nucleic acid molecules for encoding foregoing monoclonal antibody, the nucleic acid molecules have SEQ ID NO:Nucleotide sequence shown in 5-8 or the nucleotide sequence with it at least 80% homology.
Preferably, the nucleic acid molecules have SEQ ID NO:Nucleotide sequence shown in 5-8.
Encode the sequence of nucleic acid molecules such as SEQ ID NO of 1C5-F5 heavy chain of antibody variable region:Shown in 5, coding 1C5-F5 resists The sequence of nucleic acid molecules of body light chain variable region such as SEQ ID NO:Shown in 6.
Encode the sequence of nucleic acid molecules such as SEQ ID NO of 1H6-B10 heavy chain variable regions:Shown in 7,1H6-B10 light chains are encoded The sequence of nucleic acid molecules of variable region such as SEQ ID NO:Shown in 8.
The nucleic acid molecules of the foregoing monoclonal antibody of coding of the present invention include having above-mentioned preferred nucleotides sequence The nucleic acid molecules of the conserved nucleotide sequence variant of row.So-called conserved nucleotide sequence variant derived from genetic code degeneration and The variant of silence, replacement, missing and the increase of nucleotide are also included.
The present invention also provides a kind of expression vector, the expression vector includes foregoing nucleic acid molecules, in addition, also Including the expression regulation sequence being operatively connected with the sequence of nucleic acid molecules.
" carrier " word, which refers to, in the present invention can be inserted the polynucleotide for encoding certain albumen and obtain albumen A kind of nucleic acid delivery vehicle that must be expressed.Carrier can be by converting, transduceing or transfection host cell, the inhereditary material for carrying it Element is expressed in host cell.For example, carrier includes:Plasmid;Phasmid;Coemid;Artificial chromosome is such as The artificial chromosome (PAC) of yeast artificial chromosome (YAC), bacterial artificial chromosome (BAC) or P1 sources;Bacteriophage such as λ bites Thalline or M13 bacteriophages and animal virus etc..Animal virus type as carrier has retrovirus (including slow virus), gland Virus, adeno-associated virus, herpesviral (such as herpes simplex virus), poxvirus, baculoviral, papillomavirus, nipple polyoma Vacuolating virus (such as SV40).The element that a kind of carrier may be expressed containing various control, including promoter sequence, transcription initiation sequence Row, enhancer sequence, selection element and reporter gene.In addition, carrier can also contain replication origin.Carrier is it is also possible to packet It includes and its is assisted to enter ingredient of cell, such as virion, liposome or protein coat, but not only there was only these substances.
The present invention also provides a kind of host cell, foregoing nucleic acid molecules or front are contained in the host cell The expression vector.
" host cell " word refers to importing the cell of nucleic acid molecules or carrier in the present invention, including following many cells Type, such as Escherichia coli or withered grass bacterium prokaryotic cell, such as yeast cells or Aspergillus fungal cell, such as S2 drosophila cells or The insect cells such as Sf9 or such as fibroblast, Chinese hamster ovary celI, COS cells, NSO cells, HeLa cells, bhk cell, HEK The zooblast of 293 cells or people's cell.
In specific embodiments of the present invention, the host cell is Chinese hamster ovary celI.
The present invention also provides it is a kind of using foregoing host cell generate antibody method, the method includes Foregoing host cell is cultivated under the conditions of suitable and recycles the antibody.
The present invention also provides a kind of antibody generated by the above method.
The present invention also provides a kind of monoclonal antibodies for fever companion's thrombocytopenic syndromes viral glycoprotein Gc Composition, which is characterized in that the monoclonal antibody combination includes 1C5-F5 and 1H6-B10.
The present invention also provides a kind of immue quantitative detection reagent box to generate heat with thrombocytopenic syndromes viral glycoprotein, institutes It states detection kit and includes 1C5-F5 and/or 1H6-B10.
The detection kit of the present invention can only include a monoclonal antibody in 1C5-F5 or 1H6-B10, this feelings Under condition, which detects fever with thrombocytopenic syndromes virus using the association reaction of single antibody and antigen The presence of glycoprotein.
The detection kit of the present invention can include two monoclonal antibodies of 1C5-F5 and 1H6-B10 simultaneously.Such case Under, which detects fever with thrombocytopenic syndromes viral glycoprotein by double antibody sandwich ELISA In the presence of.
In specific embodiments of the present invention, 1C5-F5 is applied in combination and two kinds of monoclonal antibodies utilizations of 1H6-B10 are double Antibody sandwich ELISA method detection fever is with the presence of thrombocytopenic syndromes viral glycoprotein.
As a kind of specific embodiment, when the detection kit of the present invention utilizes 1C5-F5 and 1H6-B10 two simultaneously During kind monoclonal antibody, coated antibody is 1C5-F5;It is 1H6-B10 to detect antibody.
As a kind of embodiment that can be substituted, when the detection kit of the present invention utilizes 1C5-F5 and 1H6- simultaneously During two kinds of monoclonal antibodies of B10, coated antibody is 1H6-B10;It is 1C5-F5 to detect antibody.
Further, the foregoing detection kit of the present invention further includes ELISA Plate, confining liquid, cleaning solution, substrate colour developing Liquid, terminate liquid.
In specific embodiments of the present invention, the confining liquid is 5% milk powder.
In specific embodiments of the present invention, substrate developing solution is TMB and H2O2
In specific embodiments of the present invention, cleaning solution is phosphate buffer
In specific embodiments of the present invention, terminate liquid is the sulfuric acid of 0.5M.
A kind of detection fever the present invention also provides non-diagnostic purpose is with thrombocytopenic syndromes viral glycoprotein Method, the method detect the sugared egg of the virus using 1C5-F5 and/or 1H6-B10 by the immune response of antigen-antibody In vain.
Preferably, the method is detected using 1C5-F5 and 1H6-B10 by double antibody sandwich ELISA.
ELISA method is routine techniques well known to those skilled in the art, is repeated no more.
The present invention also provides 1C5-F5 and/or 1H6-B10 to prepare the sugared egg of companion's thrombocytopenic syndromes virus that generates heat Application in white immue quantitative detection reagent box.
The monoclonal antibody 1C5-F5 and 1H6-B10 of the present invention chemically or can pass through genetic engineering and its He is conjugated the factor.These factors provide the effect in antibody target required function site or improve or provide other property for antibody Energy.
Monoclonal antibody according to the present invention can mark chemically or by genetic engineering, detectable to provide Antibody.Detectable part includes but not limited to enzyme, prothetic group, fluorescent material, luminescent material, bioluminescent material, radioactivity Material, positron emitting metal and on-radiation paramagnetic metal ion.
In order to detect and/or analyze and/or diagnostic purpose label is dependent on particular detection/analysis/diagnostic techniques for using And/or method such as immunohistochemical staining (tissue) sample, flow cytometry, laser scanning Cytometry inspection Survey, fluorescence immunoassay, enzyme linked immunosorbent assay (ELISA) (ELISA), radiommunoassay (RIA), bioassay (such as phagocytosis make With measure), Western blotting application etc..Detection/analysis/diagnostic techniques known in the art and/or method are suitably marked It is denoted as well known to those skilled in the art.
The term as used herein " monoclonal antibody " refers to the antibody obtained from a kind of substantially uniform group, except a small number of possible Outside the existing mutation naturally occurred, the single antibody included in the group is identical.Modifier " monoclonal " only represents anti- The characteristic of body is obtained from substantially uniform antibody population, this cannot be construed to need to be produced with any specific process anti- Body.
" variable " the certain parts for representing variable region in antibody of the term as used herein are different in sequence, it is formed Combinations and specificity of the various specific antibodies to its specific antigen.Changeability, which is concentrated in light chain and heavy chain variable region, to be known as mutually It mends in three segments determined in area (CDR) or hypervariable region.Four FR areas are respectively contained in the variable region of native heavy and light chain (more conservative part in variable region), they generally in beta sheet configuration, are connected by three CDR for forming connection ring, can shape Into part β-pleated sheet structure.CDR in every chain is by FR areas firmly against together forming together and with the CDR of another chain The antigen-binding site of antibody.Constant region does not participate in the combination of antibody and antigen directly, but they show different effects Function such as participates in the cytotoxicity dependent on antibody of antibody.
The advantages of the present invention:
The detection kit of the present invention can quantify detection fever with thrombocytopenic syndromes viral glycoprotein:(1) it detects Concentration range is big;(2) high sensitivity, high specificity are reproducible;(3) available for being detected while a small amount of, multiple samples;(4) Testing cost is low;(5) detection operating procedure is few, easy to use, short the time required to detection.
Skilled person will appreciate that the fever of external preparation is with glycoprotein in thrombocytopenic syndromes viral vaccine Content is unstable, needs to be monitored the glycoprotein Content in viral vaccine, detection kit of the invention can pass through Quantitative Monitoring fever is integrated with glycoprotein Content in thrombocytopenic syndromes viral vaccine with judging to generate heat with decrease of platelet Levy the quality of viral vaccine.
Description of the drawings
Fig. 1 show using double antibody sandwich ELISA detection various concentration viral glycoprotein Gc OD450mm values it is linear Result figure.
Specific embodiment
It is further illustrated the present invention below by embodiment.It should be understood that the embodiment of the present invention is for illustrating The present invention rather than limitation of the present invention.The simple modifications that essence according to the present invention carries out the present invention belong to the present invention Claimed range.
The preparation of the grand antibody of embodiment 1 SFTSV glycoprotein (Gn and Gc) monoclonal antibody
1st, Balb/c mouse immunes
SFTSV strains (JS-2010-003) are inactivated through beta-propiolactone, low-speed centrifugal removes cell fragment, are surpassed from, molecular sieve layer Analysis technology and etc. purified.Inactivation of viruses is immunized 5 week old female Balb/c mouse (100 μ g/ only), 100 μ during first immunisation G antigens mix intraperitoneal injection with isometric Freund's complete adjuvant;It is mixed after 3rd week with equivalent amount of antigen with freund 's incomplete adjuvant Pneumoretroperitoneum is immunized;5th week the 3rd time immune, is not added with adjuvant.
2nd, splenocyte and myeloma cell are merged
It merges the last week, recovery murine myeloma cell sp2/0 to OPTI-MEM culture mediums (fetal calf serum for containing 10%), 37 DEG C are placed in, 5%CO2It cultivates in incubator, 3 days before fusion, cell is passed on primary.On the fusion same day, myeloma cell is harvested, It counts, 5.0 × 107Myeloma cell with serum free medium wash 2 times it is spare.Mouse is plucked 3-5 days after the 3rd time immune Except eyeball bloodletting, put to death.Mouse spleen is taken out in sterile working, puts in sterilizing plates, separating Morr. cell, counts, spare.
The splenocyte for being equal to 1/2 spleen of mouse is mixed with myeloma cell, 1300rpm centrifugation 5min are gone as far as possible Except supernatant.50% PEG of 1.5ml was added in 1.5 minutes, side edged shakes up;Then added in 20ml's in 8.5 minutes Serum free medium, side edged shake up.
The cell 1000rpm merged through PEG is centrifuged 5 minutes, removes supernatant, adds the HAT Selective agar medium weights of 150ml It is outstanding, fused cell is seeded to sterile 96 orifice plate, 150 μ l/ holes, is placed in 37 DEG C, 5%CO2It cultivates 4 days in incubator, is added per hole 100 μ l Selective agar mediums.
3rd, the screening of hybridoma and clone
10 days after fusion, 50 μ l supernatants are inhaled from every hole, is added to and is coated with recombination SFTSV Gn (or Gc) 96 hole ELISA Plate (is closed) with 1% BSA, is incubated at room temperature 1.0 hours;It washes 5 times.Dilute horseradish peroxidase (Horseradish Peroxidase, HRP) label goat anti-mouse 1:4000,50 μ l are added per hole, are incubated at room temperature 0.5 hour;Washing 5 times.Per hole Add in 100 μ l HRP substrates (H2O2+ TMB), it is incubated at room temperature 10 minutes, 50 μ l 0.5M H is added in per hole2SO4, survey A450nm values.
Positive hole cell is collected, is resuspended in HT Selective agar mediums, using limiting dilution assay diluting cells, and is planted in 96 It in porocyte culture plates, is observed after 5 days, to determining only to be identified as ELISA there are one the hole of cell clone growth.To sun Property hole cell carry out limiting dilution, by 3-4 time it is unicellular be separately cultured, until obtaining the hybridoma cell clone of stabilization.Greatly Amount culture hybridoma, collects the culture supernatant containing antibody, is purified with proteinG affinity columns, and carry out dialysis In PBS, survey concentration, -20 DEG C freeze it is spare.
The application double-antibody sandwich elisa selection optimum antibody pairing of embodiment 2
Obtain 7 plants of monoclonal antibodies for being directed to glycoprotein altogether by hybridoma technology, the monoclonal antibody molecule that target is Gn is 4E11-C7,3E9-B10,5D7-A6,6C5-E6, the monoclonal antibody molecule that target is Gc is 2D5-E7,1C5-F5,1H6-B10.Using Double antibody sandwich ELISA makes above-mentioned 7 strain antibody paired with each other, selects optimal combination so that glycoprotein detects, narration is such as Under:
1st, the coating of monoclonal antibody
With the NaHCO of 0.1M3/Na2CO3Buffer solution (pH 9.6) dilutes antibody concentration to 10 μ g/ml, adds in 96 hole enzymes Target, 100 μ l/ holes, 4 DEG C of coatings overnight, are washed 1 time with PBST, add in 5% 4 DEG C of milk closing overnight, with PBST wash 1 time it is standby With.
2nd, the coupling of monoclonal antibody and HRP
Monoclonal antibody is dialysed into PBS, adjusts concentration to 1mg/ml.Using the work of Beijing Thailand day and bio tech ltd Change HRP to be coupled, concrete operations are with reference to its company's specification.In labelled antibody add in 50% glycerine, -20 DEG C freeze it is standby With.
3rd, ELISA is operated
5% milk powder of 50 μ l and the inactivation of viruses culture supernatant of 50 μ l are added in per hole, 37 DEG C of incubation 1hr are washed 5 times, Then the enzyme labelled antibody (1 of 100 μ l is added in per hole:1000), 37 DEG C of incubation 0.5hr, are washed 5 times, and 100 μ l TMB are added in every hole +H2O2Substrate is incubated at room temperature 10 minutes, and the sulfuric acid that 100 μ l 0.5M are added in per hole terminates reaction, surveys A450nm values, concrete outcome As shown in table 1.
1 ELISA of table measures A450nm
Result can be seen that only 1H6-B10/1C5-F5 combinations from table, and only when 1H6-B10 is anti-as coating Body, and when 1C5-F5 is as detection antibody, the A450nm value highests of toxin are detected, reach 0.823, and other combinations A450nm values are relatively low.Above-mentioned 2 monoclonal antibodies are Gc glycoprotein specificity, therefore are combined based on 1H6-B10/1C5-F5 monoclonal antibodies Detection is Gc glycoprotein Contents in virion.
Double-antibody sandwich elisa sensitivity analysis of the embodiment 3 based on Gc glycoprotein
Using 1H6-B10 as coated antibody, 1C5-F5 and HRP conjugates as detection antibody, recombination Gc glycoprotein is target Molecule is marked, measures the susceptibility of the ELISA systems, concrete operations are same as above.
Using the detection architecture, can be detected from Virus culture supernatant down to about 60ng/ml's (being shown in Table 2 and Fig. 1) Gc antigens disclosure satisfy that SFTSV inactivated vaccines produce needs.
Table 2 is directed to the elisa assay of various concentration Gc glycoprotein
The clone of 4 monoclonal antibody 1H6-B10/1C5-F5 of embodiment weights, chain variable region gene
It takes the logarithm the hybridoma in growth period, using the Trizol extracted total RNAs of Invitrogen companies, uses oligo (dT) 20 be primer, and reverse transcription generates cDNA.Then its heavy, chain variable region gene is expanded respectively using specific primer PCR. PCR product is cloned by TA after Purified in electrophoresis and is inserted into pMD-18T carriers, and sequencing carries out sequence analysis.
The primer of amplified hybridization knurl monoclonal antibody heavy chain variable region VH is as shown in table 3.
Table 3 expands the primer sequence of heavy chain of antibody variable region
The primer sequence of amplified hybridization knurl monoclonal antibody light chain variable region VL is as shown in table 4.
Table 4 expands the primer sequence of antibody light chain variable region
As a result:
For 1H6-B10 heavy chains, only primer MHV2 (26-mer)/MHCG2b (21-mer) combinations can amplify PCR product, identified, the nucleotide sequence of 1H6-B10 heavy chain variable regions is as shown in SEQ ID NO.7, amino acid sequence such as SEQ Shown in ID NO.3.
For 1H6-B10 light chains, only primer MKV2 (30-mer)/MKC (20-mer) combinations can amplify PCR Product, identified, the nucleotide sequence of 1H6-B10 light chain variable regions is as shown in SEQ ID NO.8, amino acid sequence such as SEQ ID Shown in NO.4.
For 1C5-F5 heavy chains, only primer MHV2 (26-mer)/MHCG2b (21-mer) combinations can amplify PCR product, identified, the nucleotide sequence of 1C5-F5 heavy chain variable regions is as shown in SEQ ID NO.5, amino acid sequence such as SEQ Shown in ID NO.1.
For 1C5-F5 light chains, only primer MKV4 (33-mer)/MKC (20-mer) combinations can amplify PCR Product, identified, the nucleotide sequence of 1C5-F5 light chain variable regions is as shown in SEQ ID NO.6, amino acid sequence such as SEQ ID Shown in NO.2.
Although above only describes the specific embodiment example of the present invention, those skilled in the art should manage Solution, these are merely examples, and protection scope of the present invention is defined by the appended claims.Those skilled in the art Under the premise of without departing substantially from the principle and substance of the present invention, many changes and modifications may be made, but this A little changes or modification each fall within protection scope of the present invention.
Sequence table
<110>Jiangsu Prov. Disease Preventing and Controlling Center
<120>Fever is with thrombocytopenic syndromes viral glycoprotein immue quantitative detection reagent box
<160> 36
<170> SIPOSequenceListing 1.0
<210> 1
<211> 124
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 1
Glu Val Leu Leu Glu Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala
1 5 10 15
Ser Val Lys Ile Pro Cys Lys Thr Ser Gly Asn Gln Ile Thr Asp Tyr
20 25 30
Ser Met Asp Trp Val Lys Gln Ser His Gly Lys Thr Leu Glu Trp Ile
35 40 45
Gly Asp Ile Asn Pro Asn Asn Gly Gly Ser Ile Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Ile His Tyr Phe Gly Ser Ser Tyr Gln Pro Phe Ala
100 105 110
Tyr Trp Gly Gln Gly Thr Leu Val Ile Val Ser Ala
115 120
<210> 2
<211> 109
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 2
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Asp Ile Gly Asn Ser
20 25 30
Leu Asn Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile
35 40 45
Tyr Ala Thr Ser Ser Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly
50 55 60
Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser
65 70 75 80
Glu Asp Phe Val Asp Tyr Tyr Cys Leu Gln Tyr Ala Tyr Ser Pro Tyr
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Ala
100 105
<210> 3
<211> 118
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 3
Glu Val Leu Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Thr
1 5 10 15
Ser Val Lys Ile Pro Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30
Asn Met Asp Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp Ile
35 40 45
Gly Asp Ile Ser Pro Asn Asn Gly Gly Thr Ile Tyr Asn Gln Lys Phe
50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
65 70 75 80
Met Glu Leu Arg Ser Leu Thr Tyr Glu Asp Ser Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Tyr Gly Asn Tyr Met Arg Thr Leu Asp Tyr Trp Gly Gln Gly
100 105 110
Thr Ser Val Thr Val Ser
115
<210> 4
<211> 113
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 4
Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly
1 5 10 15
Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Gln Ser Val Ser Thr Ser
20 25 30
Ser Tyr Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
35 40 45
Lys Leu Leu Ile Lys Tyr Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60
Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Asn Ile His
65 70 75 80
Pro Val Glu Glu Glu Asp Thr Ala Thr Tyr Tyr Cys Gln His Ser Trp
85 90 95
Glu Ile Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg
100 105 110
Ala
<210> 5
<211> 373
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
gaggtcctgc tggaacagtc tggacctgag ctggtgaagc ctggggcttc agtgaagatt 60
ccctgcaaga cttctggaaa ccaaatcact gactacagca tggactgggt gaaacagagc 120
catggaaaga cccttgagtg gattggagat attaatccca acaatggtgg tagtatctac 180
aaccagaagt tcaagggcaa ggccacattg actgtagaca agtcctccag cacagcctac 240
atggagctcc gcagtctgac atctgaggac actgcagtct attactgtgc aagatacggc 300
attcattact tcggtagtag ctaccaacct tttgcttact ggggccaagg gactctggtc 360
attgtctctg cag 373
<210> 6
<211> 327
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
gacatccaga tgacccagtc tccatcctcc ttatctgcct ctctgggaga aagagtcagt 60
ctcacttgtc gggcaagtca ggacattggt aatagcttaa actggcttca gcaggaacca 120
gatggaacta ttaaacgcct gatctacgcc acatccagtt tagattctgg tgtccccaaa 180
aggttcagtg gcagtaggtc tgggtcagac tattctctca ccatcagcag ccttgagtct 240
gaagattttg tagactatta ctgtctacaa tatgcttatt ctccgtacac gttcggaggg 300
gggaccaaac tggaaataaa acgggct 327
<210> 7
<211> 354
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
gaggtcctgc tgcaacagtc tggacctgag ctggtgaagc ctgggacttc agtgaagata 60
ccctgcaagg cttctggata cacattcact gactacaaca tggactgggt gaagcagagc 120
catggaaaga gccttgagtg gattggagat attagtccta acaatggtgg tactatctac 180
aaccagaagt tcaagggcaa ggccacattg actgtagaca agtcctccag cacagcctac 240
atggagctcc gcagcctgac atatgaggac tctgcagtct attactgtgc aagatatggt 300
aattacatga ggactctgga ctactggggt caaggaacct cagtcaccgt ctcc 354
<210> 8
<211> 339
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 8
gacattgtgc tgacacagtc tcctgcttcc ttagctgtat ctctggggca gagggccacc 60
atctcatgca gggccagcca aagtgtcagt acatctagct atagttatat gcactggtac 120
caacagaaac caggacagcc gcccaaactc ctcatcaagt atgcatccaa cctagaatct 180
ggggtccctg ccaggttcag tggcagtggg tctgggacag acttcaccct caacatccat 240
cctgtggagg aggaggatac tgcaacatat tactgtcagc acagttggga gattccgtac 300
acgttcggag gggggaccaa gctggaaata aaacgggct 339
<210> 9
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 9
atgaaatgca gctggggcat sttcttc 27
<210> 10
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 10
atgggatgga gctrtatcat sytctt 26
<210> 11
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 11
atgaagwtgt ggttaaactg ggttttt 27
<210> 12
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 12
atgractttg ggytcagctt grttt 25
<210> 13
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 13
atggactcca ggctcaattt agttttcctt 30
<210> 14
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 14
atggcttgtc ytrgsgctrc tcttctgc 28
<210> 15
<211> 26
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 15
atggratgga gckggrtctt tmtctt 26
<210> 16
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 16
atgagagtgc tgattctttt gtg 23
<210> 17
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 17
atggmttggg tgtggamctt gctattcctg 30
<210> 18
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 18
atgggcagac ttacattctc attcctg 27
<210> 19
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 19
atggattttg ggctgatttt ttttattg 28
<210> 20
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 20
atgatggtgt taagtcttct gtacctg 27
<210> 21
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 21
cagtggatag acagatgggg g 21
<210> 22
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 22
cagtggatag accgatgggg c 21
<210> 23
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 23
cagtggatag actgatgggg g 21
<210> 24
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 24
caagggatag acagatgggg c 21
<210> 25
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 25
atgaagttgc ctgttaggct gttggtgctg 30
<210> 26
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 26
atggagwcag acacactcct gytatgggtg 30
<210> 27
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 27
atgagtgtgc tcactcaggt cctggsgttg 30
<210> 28
<211> 33
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 28
atgaggrccc ctgctcagwt tyttggmwtc ttg 33
<210> 29
<211> 30
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
atggatttwc aggtgcagat twtcagcttc 30
<210> 30
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
atgaggtkcy ytgytsagyt yctgrgg 27
<210> 31
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
atgggcwtca agatggagtc acakwyycwg g 31
<210> 32
<211> 31
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
atgtggggay ctktttycmm tttttcaatt g 31
<210> 33
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
atggtrtccw casctcagtt ccttg 25
<210> 34
<211> 27
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
atgtatatat gtttgttgtc tatttct 27
<210> 35
<211> 28
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 35
atggaagccc cagctcagct tctcttcc 28
<210> 36
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 36
actggatggt gggaagatgg 20

Claims (10)

1. a kind of monoclonal antibody for fever companion's thrombocytopenic syndromes viral glycoprotein Gc, which is characterized in that described Monoclonal antibody includes:
Light chain variable region, amino acid sequence such as SEQ ID NO:Shown in 2;
Heavy chain variable region, amino acid sequence such as SEQ ID NO:Shown in 1, which is named as 1C5-F5.
2. a kind of monoclonal antibody for fever companion's thrombocytopenic syndromes viral glycoprotein Gc, which is characterized in that described Monoclonal antibody includes:
Light chain variable region, amino acid sequence such as SEQ ID NO:Shown in 4;
Heavy chain variable region, amino acid sequence such as SEQ ID NO:Shown in 3, which is named as 1H6-B10.
3. encode the nucleic acid molecules of the monoclonal antibody described in claims 1 or 2, which is characterized in that the sequence of nucleic acid molecules Such as SEQ ID NO:Shown in 5-8.
4. a kind of monoclonal antibody combination for fever companion's thrombocytopenic syndromes viral glycoprotein Gc, feature exists In the monoclonal antibody combination resists including the monoclonal described in monoclonal antibody described in claim 1 and claim 2 Body.
A kind of 5. detection kit generated heat with thrombocytopenic syndromes viral glycoprotein, which is characterized in that the detection examination Monoclonal antibody of the agent box described in including monoclonal antibody described in claim 1 and/or claim 2.
6. detection kit according to claim 5, which is characterized in that the detection kit include coated antibody and/ Or detection antibody, the coated antibody is monoclonal antibody described in claim 1;The detection antibody is claim 2 institute The monoclonal antibody stated.
7. detection kit according to claim 5, which is characterized in that the detection kit include coated antibody and/ Or detection antibody, the coated antibody is the monoclonal antibody described in claim 2;The detection antibody is claim 1 institute The monoclonal antibody stated.
8. according to claim 5-7 any one of them detection kits, which is characterized in that the detection kit includes enzyme mark Plate, confining liquid, cleaning solution, substrate developing solution, terminate liquid.
9. a kind of detect method of the fever with thrombocytopenic syndromes viral glycoprotein, which is characterized in that the method includes It is immunized instead by antigen-antibody using the monoclonal antibody described in monoclonal antibody described in claim 1 and/or claim 2 It should detect, it is preferable that passed through using the monoclonal antibody described in monoclonal antibody described in claim 1 and claim 2 Double antibody sandwich ELISA detects.
10. the monoclonal antibody described in claims 1 or 2 is preparing companion's thrombocytopenic syndromes viral glycoprotein detection of generating heat Application in kit.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108956588A (en) * 2018-07-09 2018-12-07 东南大学 Electrochemical luminescence immunosensor is used to prepare serious fever with the application of decrease of platelet syndrome virus detection kit
CN110437332A (en) * 2019-08-20 2019-11-12 江苏省疾病预防控制中心(江苏省公共卫生研究院) A kind of SFTSV protein binding molecule of viral infection resisting
CN110684102A (en) * 2019-07-23 2020-01-14 源道隆(苏州)医学科技有限公司 SFTSV detection kit
CN112342316A (en) * 2020-10-28 2021-02-09 中国疾病预防控制中心病毒病预防控制所 Primer probe set and detection method for detecting fever with thrombocytopenia syndrome virus by real-time fluorescent RNA isothermal rapid amplification

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102809653A (en) * 2012-04-24 2012-12-05 万里明 Preparation and application of ELISA (Enzyme-Linked Immunosorbent Assay) kit for detecting novel bunyavirus antigen
CN102942629A (en) * 2012-11-21 2013-02-27 江苏省疾病预防控制中心 Humanized antibody for resisting severe fever with thrombocytopenia syndrome bunyavirus (SFTSV)

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102809653A (en) * 2012-04-24 2012-12-05 万里明 Preparation and application of ELISA (Enzyme-Linked Immunosorbent Assay) kit for detecting novel bunyavirus antigen
CN102942629A (en) * 2012-11-21 2013-02-27 江苏省疾病预防控制中心 Humanized antibody for resisting severe fever with thrombocytopenia syndrome bunyavirus (SFTSV)

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
李阿茜等: "《抗发热伴血小板减少综合征病毒结构蛋白单克隆抗体的制备和功能分析》", 《病毒学报》 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108956588A (en) * 2018-07-09 2018-12-07 东南大学 Electrochemical luminescence immunosensor is used to prepare serious fever with the application of decrease of platelet syndrome virus detection kit
CN110684102A (en) * 2019-07-23 2020-01-14 源道隆(苏州)医学科技有限公司 SFTSV detection kit
CN110713536A (en) * 2019-07-23 2020-01-21 源道隆(苏州)医学科技有限公司 Polypeptide capable of combining SFTSV, nucleic acid coding sequence and application thereof
CN110684102B (en) * 2019-07-23 2022-10-21 源道隆(苏州)医学科技有限公司 SFTSV detection kit
CN110437332A (en) * 2019-08-20 2019-11-12 江苏省疾病预防控制中心(江苏省公共卫生研究院) A kind of SFTSV protein binding molecule of viral infection resisting
CN110437332B (en) * 2019-08-20 2020-03-06 江苏省疾病预防控制中心(江苏省公共卫生研究院) SFTSV protein binding molecule for resisting virus infection
CN112342316A (en) * 2020-10-28 2021-02-09 中国疾病预防控制中心病毒病预防控制所 Primer probe set and detection method for detecting fever with thrombocytopenia syndrome virus by real-time fluorescent RNA isothermal rapid amplification

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