CN101333248A - B cell antigen epitope polypeptide of encephalitis B virus E protein and uses thereof - Google Patents
B cell antigen epitope polypeptide of encephalitis B virus E protein and uses thereof Download PDFInfo
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Abstract
The invention discloses a polypeptides epitope in Japanese encephalitis virus E protein neutralizing B cell, also discloses the application of the polypeptides epitope in the prevention, treatment and diagnosis of Japanese encephalitis, belonging to the field of molecular immunology. The amino acid sequence of the polypeptides epitope disclosed in the invention is selected from any of the amino acid shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 5. After being used as immunogen or vaccine immune animal organisms, the JEV E protein neutralizing B cell polypeptides epitope of the invention can produce neutralizing antibody against Japanese encephalitis virus, and can neutralize the Japanese encephalitis virus in vivo or in vitro so as to prevent the virus infecting animal organisms. The polypeptides epitope of the invention or the conjugate of the polypeptides epitope can be used as the reagents for detecting the anti-Japanese encephalitis virus antibodies or anti-Japanese encephalitis virus polypeptide antibodies.
Description
Technical field
The present invention relates to antigen epitope polypeptide, relate in particular to encephalitis b virus E albumen neutrality B cell antigen epitope polypeptide, the invention still further relates to the application of this antigen epitope polypeptide in control and diagnosis encephalitis b virus, belong to the molecular immunology field.
Background technology
The epidemic encephalitis type B encephalitis B is that (Japanese Encephalitis virus, a kind of important mosquito matchmaker property people beast that JEV) causes suffers from transmissible disease altogether by encephalitis b virus.This virus is at first separated acquisition in nineteen fifty-three in Japan from patient's cerebral tissue, therefore claim japanese encephalitis virus, and associated diseases claims Japanese B encephalitis in Japan.Separated this virus in China,, name and be epidemic encephalitis type B, be called for short encephalitis in order to distinguish mutually with encephalitis A early than 1940.Worldwide have 30 every year, 000-50,000 routine encephalitis case, about 25%-30% death, 50% causes permanent central nervous system sequela.The annual in recent years encephalitis case of China is greater than 5000 examples, more than dead 200 examples.For the multiple animal susceptible of epidemic encephalitis B virus, but except that people, horse and pig, do not present clinical symptom usually.The people is mainly caused the acute infection of central nervous system, and it is feature with high heat, the disturbance of consciousness, tic, respiratory insufficiency and meningeal irritation sign clinically that the people infects.Sow infects can cause miscarriage, produce stillborn foetus or mummy tire, and it is feature that the boar infection causes the testis enlargement to influence reproductive performance, and the minority pig only infects and may present heating and nervous symptoms, and other pigs then great majority do not manifest symptom.Cause enormous economic loss to pig industry.Equus easy infection, minority cause encephalitis and nervous symptoms, and horse is the final host.Mosquito is the main communication media of JEV, and pig is that this virus is most important storage of occurring in nature and propagation host.Encephalitis b virus is mainly popular in the Asia, and recent findings also has report in the Southern Hemisphere, and this explanation epidemic encephalitis type B may worldwide threaten human health.
Encephalitis b virus belongs to flaviviridae, and Flavivirus is the strand positive chain RNA virus.The about 11kb of genome comprises a big open reading frame (ORF), and two ends are 5 ' end and 3 ' end non-coding region (UTR).Genome 5 ' end has cap sequence, but does not have poly (A) tail at 3 ' end.ORF coding contains 3432 amino acid whose precursor proteins, is processed as 10 maturation proteins under the effect of virus and host cell proteins enzyme, comprise 3 structural protein (C, prM, E), 7 Nonstructural Proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).Virus particle is spherical in shape, and the about 30nm of diameter has cyst membrane.JEV has only a serotype, belongs to same serogroups with western sieve virus, black Luo Hegu encephalitis, St. Louis encephalitis virus etc. on antigenicity, has antigenic cross-reaction.Virus envelope glycoprotein has the blood clotting characteristic, animal erythrocytes such as energy aggegation goose, dove, chick.Virus is to a little less than the heat resist power, and 56 ℃ of 30 minutes or 100 ℃ got final product deactivation in 2 minutes, but very strong to low temperature and exsiccant resistibility.Ether, 1: but the equal inactivation of viruses of 1000 sodium deoxycholate and common disinfectants.Unstable under acidic condition, appropriate pH 8.5～9.0.
Adopting vaccine to carry out preventive vaccination is these disease popular effective measure of control.To the research of encephalitis b virus epitope is exploitation effective gene engineered vaccine and the research theoretical basis based on the diagnostic detect reagent of epitope.E albumen is a kind of important structural protein of encephalitis b virus particle surface, is important immune protective antigen.The B cell antigen epi-position that has neutralizing effect in protective antigen plays and must act in stimulating body generation neutrality antibody.Identify that correctly and at length artificial vaccine that epitope has no side effect to the diagnosis of disease, design and immunotherapeutic agent etc. have important meaning.
Summary of the invention
One of purpose of the present invention provides encephalitis b virus JEV E protein B cell antigen epitope polypeptide.
Two of purpose of the present invention is that described encephalitis b virus JEV E protein B cell antigen epitope polypeptide is applied to treatment or prevention encephalitis b virus, perhaps uses it for the reagent that is prepared into diagnosis or detects encephalitis b virus.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A kind of encephalitis b virus JEV E protein B cell antigen epitope polypeptide, its aminoacid sequence is selected from any one aminoacid sequence shown in SEQ IDNO:1, SEQ ID NO:2, SEQ ID NO:3 or the SEQ ID NO:4.
Wherein, can carry out chemically modified to the N end or the C end of described encephalitis b virus JEV E protein B cell antigen epitope.Described chemically modified is the natural amination or the acetylize of polypeptide chain N end, or the carboxylated naturally or amidation of C end.
Encephalitis b virus JEV E protein B cell antigen epitope described in the present invention can obtain by solid-phase peptide synthesis is synthetic, also can prepare by genetic engineering technique, and these are all understood thoroughly by those skilled in the art.
Encephalitis b virus JEV E protein B cell antigen epitope of the present invention or its connector can be used for preparing immunogen, or with its immune animal to produce immunoprotection.Also described encephalitis b virus JEV E protein B cell antigen epitope or its connector can be prepared into the reagent that detects anti-encephalitis b virus JEV antibody or anti-encephalitis b virus JEV polypeptide antibody.
Epitope polypeptide of the present invention is prepared into immunogen or vaccine, at least comprise described one or whole epitope, these polypeptide connect, interconnect or are connected with carrier by self, and are aided with the T cell antigen epitope, comprise Th1 and/or Th2 epitope polypeptide.These T cell antigen epitopes can derive from has the immunocompetent peptide sequence of the body cell of stimulation in JEV viral protein or other animal proteinum.
Detecting encephalitis b virus JEV can be at the monoclonal antibody of E proteantigen epi-position or at the polyclonal antibody of antigen epitope polypeptide, detection method can comprise indirect ELISA, double-antibody sandwich elisa, direct immunofluorescence technic, IiT, and quick detection test paper bar immunity rete is analysed technology etc.
Detect encephalitis b virus JEV antibody, comprise that wild virus infection produces antibody, vaccine immunity produces antibody, maternal antibody, anti-JEV monoclonal antibody etc.Detection method can comprise indirect ELISA, double-antibody sandwich elisa, and quick detection test paper bar immunity rete is analysed technology etc.
The method that proteantigen B cell antigen epi-position is identified has multiple, as genetically engineered rite-directed mutagenesis antigen expressed analysis of protein method, phage random peptide library triage techniques, synthetic polypeptide detection technique etc.What this invention mainly adopted in identifying epitope is that the genetic engineering technique polypeptide merges overlapping (overlapping) expression JEV E albumen, external immune response detects the polypeptide amalgamation protein group and the sero-fast associativity of JEV is identified the epitope sequence, further shorten and mutation expression epi-position fusion rotein, identify the sequential structure of epitope; Use the exactness of synthetic polypeptide technical identification epi-position again; Prepare epitope specificity single-factor serum with epi-position fusion rotein antigen-immunized animal, analyze the epitope function.
Beneficial effect of the present invention is as follows:
1. by the antibody that can produce behind the immunogen of encephalitis b virus JEV E protein B cell antigen epitope design or the vaccine immunity animal body, utilize monoclonal antibody or polyclonal antibody can detect encephalitis b virus JEV or encephalitis b virus E albumen at E proteantigen epi-position at JEV.
2. encephalitis b virus E protein B cell antigen epitope of the present invention is connected with carrier or self connects or interconnects; can immune animal, anti-peptide antibody that behind immune animal, produces or anti-B encephalitis virus antibody can be in vivo and in vitro in and encephalitis b virus JEV and produce immunoprotection.
3. encephalitis b virus E protein B cell antigen epitope of the present invention or its connector can detect encephalitis b virus JEV antibody or anti-encephalitis b virus JEV polypeptide antibody.
Description of drawings
Fig. 1 is epitope polypeptide SEQ ID NO:1 to SEQ ID NO:4 and GST amalgamation and expression albumen and anti-JEV serum Western blot detected result.Be followed successively by from left to right among the figure: M, protein molecular weight standard; 1, GST-SEQ ID NO:1; 2, GST-SEQ ID NO:2; 3, GST-SEQ ID NO:3; 4, GST-SEQID NO:4; 5, the GST contrast.
Fig. 2 be epitope polypeptide antibody external in and the encephalitis b virus experimental result.1, the negative serum contrast; 2, anti-epitope polypeptide Seq No 3 serum.
Embodiment
According to JEV E Argine Monohydrochloride sequence, design one adds up to 36 serial polypeptide, these polypeptide are overlapped, and cover JEV E albumen 1-296 amino-acid residue zone, and this zone comprises E protein structure domain I (Domain I) and domain II (Domain II).According to each bar polypeptid acid sequence, the synthetic a pair of DNA chain of design inserts expression vector pGEX-6p-1 with the DNA chain and GST carries out amalgamation and expression.The serial recombinant protein of amalgamation and expression and anti-JEV serum carry out the indirect ELISA analysis, analyze JEV E proteantigen epi-position.Idiographic flow is as follows:
DNA chain → polypeptide fusion expression vector structure → abduction delivering → expressed fusion protein affinitive layer purification → the expression product of design peptide sequence → composite coding peptide sequence and the reactive detection → analysis JEVE of anti-JEV seroimmunity proteantigen epi-position.
Table 1 polypeptide amalgamation protein and anti-JEV serum indirect ELISA reaction result
Annotate: envelope antigen is respectively polypeptide amalgamation protein, and GST is the contrast envelope antigen, and antigen coated concentration is 10 μ g/ml; Anti-JEV serum is 200 times of dilutions.
Synthesizing of embodiment 2 encephalitis b virus JEV E protein B cell antigen epitopes
According to the aminoacid sequence of JEV E protein B cell antigen epitope, use the solid-phase polypeptide synthetic technology, adopt automatic Peptide synthesizer or artificial polypeptide synthesis method.Solid-phase resin adopts amino acid Wang resin or other resin of Fmoc protection, and resin according to the aminoacid sequence order, adds the amino acid of Fmoc protection after DMF swelling, piperidine glue Fmoc protection, exist at HBTU and carry out acylation reaction under the situation.Through washing, add deputy Fmoc-amino acid again and carry out acylation reaction after acylation reaction is finished, and washing.So circulation is super initial to N-terminal from the peptide sequence C-terminal, synthetic in order complete polypeptide chain.After synthetic the finishing, choose suitable reagent with being connected and precipitating the TFA polypeptide of TFA method cracking peptide chain and resin with cold diethyl ether according to component peptide chain amino acid difference, standby after desalting and purifying, LC-MS and HPLC Analysis and Identification.In entire synthesis process, can use the performance of Kaiser method or TMBS method test reaction after each amino acid acylation reaction.For the ease of epitope polypeptide is connected with carrier proteins, when synthesizing polypeptide, can add a halfcystine at the N-terminal of peptide sequence.
According to the method described above, the polypeptide fragment that has synthesized aminoacid sequence shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and the SEQ ID NO:4 respectively.
The chemically modified of embodiment 3 encephalitis b virus JEV E protein B cell antigen epitopes
According to the method synthetic polypeptide fragment of embodiment 3, its N-terminal is that nature is amido modified, and C-terminal is the nature carboxyl modified.
N-terminal acetylation modification method: the polypeptide according to above introduction is blended into last Fmoc protection amino acid coupling end, and carries out the removal of N-terminal Fmoc blocking group, carries out following operation then.Get 150 μ l diacetyl oxides and 20 μ L EIPEA are dissolved among the 4.8ml DMF, thorough mixing also cools off in ice bath.Then above mixed solution is added in the polypeptide building-up reactions pipe and react 5min.Use 5ml DMF and methylene dichloride (DCM) to wash again successively 2 times, each 5min.Nitrogen dries up the back and carries out removal, cracking, purifying and the evaluation of side chain protected group according to ordinary method.
C-terminal amidation modifying method: should select resin for use when the amidated polypeptide of C-terminal is synthetic is the Rink resin.The reply resin begins to carry out synthesis program from C-terminal first amino acids then with DMF swelling 20min when carrying out synthesis program, the same Fmoc-wang resin synthetic method of crossing.
The coupling of embodiment 4 encephalitis B JEV E protein B cell antigen epitopes and carrier proteins KLH and BSA:
Carrier proteins KLH or BSA 4mg are dissolved in 0, and 5ml contains in PBS (pH7.2) damping fluid of 5mM EDTA, add 1.0mg linking agent Sulfo-SMCC, and fully 30min is hatched incubated at room 60min or 37 ℃ in the dissolving back.Use the carrier proteins that Sephadex G-25 post or dialysis process desalting and purifying Sulfo-SMCC handle, and with standby behind the BCA method mensuration protein content.
Take by weighing synthetic epitope polypeptide 4mg, add 0.5ml double distilled water dissolving polypeptide, the polypeptide after will dissolving then mixes with the carrier proteins that Sulfo-SMCC handles, and hatches 2 hours or reacts and spend the night for 4 ℃, and packing is standby.
The polypeptide that contains halfcystine can form self and connect by the oxidizing reaction of sulfydryl.The oxidizing reaction of using the DMSO mediation generally speaking realizes connecting, and according to the acid-basicity of polypeptide, oxidizing reaction can be carried out under little acid or slight alkalinity condition.
Oxidizing reaction under the slightly acidic environment: with the acetate dissolution polypeptide of proper concn, the concentration that makes polypeptide is in the 0.5-1.5mM scope, the final concentration of acetic acid is no more than 5%, the pH that adjusts polypeptide solution with (NH4) 2CO3 is about 6.0, the DMSO that adds 10-20%, 25 ℃ of reaction 5-25h, the carrying out degree and can monitor of oxidizing reaction by HPLC.Then, be moving phase with 1%TFA water and acetonitrile, preparation type reversed-phase HPLC purifying oxidisability polypeptide.
Oxidizing reaction under the slight alkalinity environment: use the 0.01M phosphate buffered saline buffer, pH7.5, dissolving polypeptide add concentration DMSO and spend the night to 1%, 25 ℃ of reaction of final concentration to final concentration 1.0mM, the carrying out degree and can monitor by HPLC of oxidizing reaction.Be moving phase with 1%TFA water and acetonitrile then, preparation type reversed-phase HPLC purifying oxidisability polypeptide.
Embodiment 6 encephalitis b virus JEV E protein B cell antigen epitope indirect ELISAs detect the anti-JEV antibody of pig
Synthesizing the polypeptide or the connector of epitope polypeptide and carrier or the mixture of epitope polypeptide self connector with JEV E protein B cell antigen epitope SEQ ID NO:1 to SEQ ID NO:4 is envelope antigen, burden 96 hole polystyrene enzyme plates.Use pH9.6,0.1M carbonate buffer solution dilution antigen to final concentration is 10 μ g/ml, adds in the enzyme plate by 100 μ l/ holes, and 4 ℃ of bags are spent the night.Use PBST (PBS+0.05%Tween) detersive enzyme target 3 times then; The PBST sealase target that contains 1%BSA, 37 ℃ of sealing 2h, plate is washed 3 times with washing lotion PBST in sealing back, is used for detection or-20 ℃ immediately and deposits standby.
Antibody test schedule of operation: add porcine blood serum to be detected (100 times of dilutions of qualitative detection serum, antibody titer detects serum and carries out doubling dilution, set up positive serum contrast and negative serum contrast and the blank of increase serum not simultaneously), hatch 1h for 37 ℃, PBST washing lotion washing 3 times, each 3 minutes; Add horseradish peroxidase-labeled goat-anti pig IgG (Goat-anti-pig IgG-HRP), hatch 1h for 37 ℃, PBST washing lotion washing 4 times, each 3 minutes; Add HRP chromogenic substrate (TMB developer), incubated at room 5-15 minute observation color reaction; Fully after the colour developing, add the reaction of 2M sulfuric acid color development stopping; Measure the light absorption value of 450nm wavelength with microplate reader; Result of determination.
During result of determination: blank and negative serum hole light absorption value are less than or equal to 0.2, and positive serum control wells light absorption value is effective greater than 0.4 o'clock result; Calculate P/N value=(detection hole OD value-blank hole OD value)/(negative serum OD value-blank hole OD value), the P/N value be equal to or greater than 2 o'clock positive; Maximum dilution multiple with reacting positive serum is the antibody titer of this sample serum.
Table 2 mixed polypeptide bag is detected the anti-JEV antibody of pig by indirect ELISA
Annotate: envelope antigen is the mixture of the synthetic polypeptide of epitope SEQ ID NO:1 to SEQ ID NO:4,10 μ g/ml.It is 200 that this positive serum is tired.
Embodiment 7 encephalitis b virus JEV E protein B cell antigen epitope indirect ELISAs detect the anti-JEV antibody of people
Implementation method is with embodiment 6, and serum to be checked, negative control sera and positive control serum are human serum; Two anti-are horseradish peroxidase-labeled goat anti-human igg (Goat-antihuman IgG-HRP).
Embodiment 8 encephalitis b virus JEV E protein B cell antigen epitope indirect ELISAs detect the anti-JEV antibody of horse
Implementation method is with embodiment 6, and serum to be checked, negative control sera and positive control serum are horse serum; Two anti-are horseradish peroxidase-labeled goat-anti horse IgG (Goat-antihorse IgG-HRP).
Embodiment 9 encephalitis b virus JEV E protein B cell antigen epitope Dot-ELISA detect the anti-JEV antibody of pig
The mixture of the connector of JEV E protein B cell antigen epitope SEQ ID NO:1 to SEQ ID NO:4 polypeptide or epitope polypeptide and carrier is an envelope antigen, is contrast antigen with BSA.The envelope antigen of on nitrocellulose filter, fixing a point, 2 μ g/ points, 37 ℃ of fixing 30min, PBST washing 6 times, 4 ℃ of sealings are spent the night among the 2%BSA, promptly can be used for antibody test after the PBST washing 6 times.
When detecting antibody, serum sample to be detected is suitably diluted the back hatch 30min, PBST washing 6 times, adding horseradish peroxidase-labeled goat-anti pig IgG (Goat-anti-pig IgG-HRP) in 37 ℃ with antigen coated film, hatch 1h for 37 ℃, PBST washing lotion washing 6 times; Add the colour developing of AEC or DAB substrate, the room temperature lucifuge is hatched 10min observation color reaction.
When the result judges, present the red-brown color reaction with the envelope antigen point, contrast antigen point color reaction does not the occur, and the person is positive.
It is anti-that embodiment 10 encephalitis b virus JEV E protein B cell antigen epitope Dot-ELISA detect the anti-JEV of people
Implementation method is with embodiment 9, and serum to be checked is human serum; Two anti-are horseradish peroxidase-labeled goat anti-human igg (Goat-anti human IgG-HRP).
It is anti-that embodiment 11 encephalitis b virus JEVE protein B cell antigen epitope Dot-ELISA detect the anti-JEV of horse
Implementation method is with embodiment 9, and serum to be checked is horse serum; Two anti-are horseradish peroxidase-labeled goat-anti horse IgG (Goat-anti horse IgG-HRP).
Embodiment 12 immunogenic preparation of encephalitis b virus JEV E protein B cell antigen epitope and immunization experiments
Encephalitis b virus JEV E protein B cell antigen epitope immunogen can be the synthetic antigen epitope polypeptide, antigen epitope polypeptide self connector, and antigen epitope polypeptide and carrier connector, epitope merges epi-position albumen etc.With antigen epitope polypeptide SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:4 and BSA connector are immunogen in the present embodiment; First immunisation immunogen Freund's complete adjuvant and antigen emulsification, animalcule immunizing antigen amount is every of 100 μ g; Two or three when exempting from Freund's incomplete adjuvant and antigen emulsification, animalcule immunizing antigen amount is that every of 50-100 μ g carries out immunity.Immunization route can be subcutaneous injection, intradermal injection or intramuscular injection.Each time duration of immunity is two weeks at interval.Antibody horizontal is detected in twice back of booster immunization.
Test-results sees Table 3 respectively, table 4 and table 5.
Table 3 mouse anti epitope polypeptide SEQ NO:1 antibody indirect ELISA is tired
Annotate: envelope antigen is antigen epitope polypeptide SEQ NO:1,10 μ g/ml.
Table 4 mouse anti epitope polypeptide SEQ NO:3 antibody indirect ELISA is tired
Annotate: envelope antigen is antigen epitope polypeptide SEQ NO:3,10 μ g/ml.
Table 5 mouse anti epitope polypeptide SEQ NO:4 antibody indirect ELISA is tired
Annotate: envelope antigen is antigen epitope polypeptide SEQ NO:4,10 μ g/ml.
Sequence table
＜110〉Harbin Veterinary Medicine Inst., China Academy of Agriculture
＜120〉B cell antigen epitope polypeptide of encephalitis B virus E protein and application thereof
<130>KLPI012
<160>4
<170>PatentIn?version?3.5
<210>1
<211>16
<212>PRT
<213>Japanese?Encephalitis?virus
<400>1
Phe?Asn?Cys?Leu?Gly?Met?Gly?Asn?Arg?Asp?Phe?Ile?Glu?Gly?Ala?Ser
1 5 10 15
<210>2
<211>12
<212>PRT
<213>Japanese?Encephalitis?virus
<400>2
Glu?Lys?Arg?Ala?Asp?Ser?Ser?Tyr?Val?Cys?Lys?Gln
1 5 10
<210>3
<211>16
<212>PRT
<213>Japanese?Encephalitis?virus
<400>3
Gly?Thr?Thr?Thr?Ser?Glu?Asn?His?Gly?Asn?Tyr?Ser?Ala?Gln?Val?Gly
1 5 10 15
<210>4
<211>16
<212>PRT
<213>Japanese?Encephalitis?virus
<400>4
Ser?Gln?Glu?Gly?Gly?Leu?His?His?Ala?Leu?Ala?Gly?Ala?Ile?Val?Val
1 5 10 15
Claims (7)
1, a kind of encephalitis b virus JEV E protein B cell antigen epitope polypeptide, it is characterized in that: its aminoacid sequence is selected from wherein any one aminoacid sequence shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 or the SEQ ID NO:4.
2, the nucleotide sequence of the described encephalitis b virus JEV of coding claim 1 E protein B cell antigen epitope polypeptide.
3, encephalitis b virus JEV E albumen neutrality B cell antigen epitope polypeptide according to claim 1 is characterized in that: described encephalitis b virus JEV E albumen neutrality B cell antigen epitope polypeptide carries out chemically modified at N end or C end.
4, encephalitis b virus JEV E albumen neutrality B cell antigen epitope polypeptide according to claim 4 is characterized in that: the chemically modified of described polypeptide chain N end is the natural amination or the acetylize of N end; The chemical modification of described polypeptide chain C end is the carboxylated naturally or amidation of C end.
5, the vaccine composition of a kind of prevention or treatment encephalitis b virus is characterized in that: contain the described encephalitis b virus E of the claim 1 albumen neutrality B cell antigen epitope polypeptide that significant quantity is gone up in treatment.
6, the purposes of the described encephalitis b virus E of claim 1 albumen neutrality B cell antigen epitope polypeptide in preparation prevention or treatment encephalitis b virus medicine.
7, the purposes of the described encephalitis b virus E of claim 1 albumen neutrality B cell antigen epitope polypeptide in preparation diagnosis or detection encephalitis b virus and antibody reagent thereof.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101492495B (en) * | 2009-02-24 | 2011-08-31 | 中国农业科学院哈尔滨兽医研究所 | A group of antigen epitope polypeptide and uses thereof |
| CN102206249A (en) * | 2011-04-26 | 2011-10-05 | 中国农业科学院哈尔滨兽医研究所 | Specific B cell epitope polypeptide of NS1 protein of encephalitis B virus and use thereof |
| CN101607983B (en) * | 2009-07-29 | 2012-06-13 | 中国农业科学院哈尔滨兽医研究所 | Encephalitis B virus PrM/M protein B cell antigen epitope polypeptide and applications |
| CN110709410A (en) * | 2017-04-26 | 2020-01-17 | 豪夫迈·罗氏有限公司 | Soluble and immunoreactive Zika virus NS1 polypeptide |
| CN114621343A (en) * | 2022-05-17 | 2022-06-14 | 首都医科大学 | Japanese encephalitis virus antibody 2G1 and application thereof |
-
2008
- 2008-07-30 CN CNA2008101262909A patent/CN101333248A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101492495B (en) * | 2009-02-24 | 2011-08-31 | 中国农业科学院哈尔滨兽医研究所 | A group of antigen epitope polypeptide and uses thereof |
| CN101607983B (en) * | 2009-07-29 | 2012-06-13 | 中国农业科学院哈尔滨兽医研究所 | Encephalitis B virus PrM/M protein B cell antigen epitope polypeptide and applications |
| CN102206249A (en) * | 2011-04-26 | 2011-10-05 | 中国农业科学院哈尔滨兽医研究所 | Specific B cell epitope polypeptide of NS1 protein of encephalitis B virus and use thereof |
| CN102206249B (en) * | 2011-04-26 | 2013-07-03 | 中国农业科学院哈尔滨兽医研究所 | Specific B cell epitope polypeptide of NS1 protein of encephalitis B virus and use thereof |
| CN110709410A (en) * | 2017-04-26 | 2020-01-17 | 豪夫迈·罗氏有限公司 | Soluble and immunoreactive Zika virus NS1 polypeptide |
| CN114621343A (en) * | 2022-05-17 | 2022-06-14 | 首都医科大学 | Japanese encephalitis virus antibody 2G1 and application thereof |
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