CN103936852A - Nanometer antibody for specifically aiming at H3N2 influenza A virus and application thereof in diagnosis - Google Patents
Nanometer antibody for specifically aiming at H3N2 influenza A virus and application thereof in diagnosis Download PDFInfo
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- CN103936852A CN103936852A CN201410202193.9A CN201410202193A CN103936852A CN 103936852 A CN103936852 A CN 103936852A CN 201410202193 A CN201410202193 A CN 201410202193A CN 103936852 A CN103936852 A CN 103936852A
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Abstract
The invention discloses a nanometer antibody for specifically aiming at an H3N2 influenza A virus, a gene sequence for encoding the nanometer antibody and an application of the nanometer antibody in diagnosis, solving the problem that the production and detection of a high-purity high-affinity high-sensitivity antibody can not be obtained in the research process of the traditional H3N2 influenza A diagnostic reagent. The nanometer antibody for specifically aiming at the H3N2 influenza A virus, which is disclosed by the invention, can be used for fast detecting the antigen by being efficiently expressed in escherichia coli and being improved and is applied to the research of an H3N2 influenza A fast diagnosis kit.
Description
Technical field
The invention belongs to biomedicine or biological pharmacy technical field, relate to nano antibody, nano antibody renovation technique and the research and development for quick diagnosis reagent kit thereof of specificity for A type H3N2 influenza virus.
Background technology
The difference of influenza A virus hemagglutinin (HA) antigen surperficial according to it can be divided into 16 hypotypes (H1-H16), can be divided into 9 hypotypes (N1-N9) according to the difference of neuraminidase (NA) antigen.The accumulation of influenza virus by surface protein antigen changes (antigenic drift) or various flows Influenza Virus producer and resets the immunity identification that produces a kind of new influenza virus (antigenic shift) and escape infection host, thus infect in colony propagate, procreation.A type H3N2 influenza is a kind of respiratory system disease causing because of A type H3N2 influenza virus.The H3N2 influenza of nineteen sixty-eight Hong Kong outburst is to million people is lost one's life.At the beginning of 2013, the multiple state outburst in eastern united states H3N2 seasonal influenza, classic city government announced the public health emergency state January 9.At present mainly adopt chemicals and vaccine for the prevention and control measure of human world A type H3N2 flu outbreak, but chemicals excessive use the raw side effect of fecund and vaccine development often lag behind epidemic situation break out can only the touching prophylactic effect of all rising of commute with vaccine drawback remarkable.And having manifested good effect by antibody-mediated prevention and diagnosis, its application prospect obtains expert's approval.But conventional antibody still existence and stability is poor, solvability is low and obtain the problem of the aspect such as trouble.The macromole characteristic of antibody drug has restricted it and has been widely used and the performance of curative effect.The digested system of general antibody capable is degraded very soon, thereby has stoped its periphery that enters brain or some tumours, and numerous disease cannot be treated with monoclonal antibody drug.
And nano antibody is at present minimum antibody molecule, its molecular weight is 1/10 of common antibody, and at first by Belgian scientist Hamers, R finds in camel blood, and it is the class receiving much concern in engineered antibody product.It has the unique advantages such as molecule is little, stability is strong, solubility is good, immunogenicity is low, easy expression, easy transformation, and can need to carry out personalized designs easily to it according to what apply, facilitating other molecules of coupling, is a large focus of current antibody product exploitation.Nano antibody has developed into has extensive biologic applications to be worth and the general molecular of clinical value, and its application relates to multiple fields such as medical diagnosis on disease and treatment, basic medical research, biological study.Utilize nano antibody technology to obtain the nano antibody of two strains for the different epi-positions of A type H3N2 influenza virus, it is carried out to improvement and design, for the diagnostic kit research and development of rapid detection antigen.
Summary of the invention
Goal of the invention: technical problem to be solved by this invention is to provide the nano antibody of two strains for the different epi-positions of A type H3N2 influenza virus provides the application of this nano antibody in diagnostic kit research and development simultaneously.
Technical scheme: for achieving the above object, a first aspect of the present invention, a kind of VHH chain of H3N2 nano antibody is provided, comprise framework region FR and complementary determining region CDR, described framework region FR is made up of four framework regions, be respectively FRl, FR2, FR3 and FR4, their aminoacid sequence is selected from following aminoacid sequence:
FR1 is the aminoacid sequence shown in SEQ ID NO.1, and FR2 is the aminoacid sequence shown in SEQ ID NO.2, and FR3 is the aminoacid sequence shown in SEQ ID NO.3, and FR4 is the aminoacid sequence shown in SEQ ID NO.4;
Or FR1 is the aminoacid sequence shown in SEQ ID NO.5, FR2 is the aminoacid sequence shown in SEQ ID NO.6, and FR3 is the aminoacid sequence shown in SEQ ID NO.7, and FR4 is the aminoacid sequence shown in SEQ ID NO.8;
Described complementary determining region CDR is made up of three complementary determining regions, CDRl, CDR2 and CDR3 respectively, and their aminoacid sequence is selected from following aminoacid sequence:
CDR1 is the aminoacid sequence shown in SEQ ID NO.9, and CDR2 is the aminoacid sequence shown in SEQ ID NO.10, and CDR3 is the aminoacid sequence shown in SEQ ID NO.11;
Or CDR1 is the aminoacid sequence shown in SEQ ID NO.12, CDR2 is the aminoacid sequence shown in SEQ ID NO.13, and CDR3 is the aminoacid sequence shown in SEQ ID NO.14.
Preferably, described specificity is for the nano antibody of A type H3N2 influenza virus, and it has the aminoacid sequence shown in SEQ ID NO.15 or 16.
Second aspect present invention, a kind of H3N2 nano antibody, it is the nano antibody of specificity for A type H3N2 influenza virus epi-position, comprises two VHH chains with aminoacid sequence shown in SEQ ID NO.15 or 16.
Third aspect present invention, a kind of DNA molecular, its coding is selected from the protein of lower group: the VHH chain of the H3N2 nano antibody described in claim 1 or 2, or H3N2 nano antibody claimed in claim 3.
Preferably, described DNA molecular, it has the DNA sequence dna that is selected from lower group: the aminoacid sequence shown in SEQ ID NO.17 or 18.
A fourth aspect of the present invention, a kind of expression vector, it is containing the nucleotide sequence shown in SEQ ID NO.17 or 18.
A fifth aspect of the present invention, a kind of host cell, it can express the nano antibody for A type H3N2 influenza virus.
H3N2 nano antibody external source is added vitamin H and coupling horseradish peroxidase, and is applied to the research and development of quick detection kit.
Beneficial effect: compared with prior art, advantage of the present invention is as follows: the present invention utilizes the A type H3N2 influenza virus natural antigen immunity camel of deactivation, obtains high-quality immune nano library of antibody genes.Then the A type H3N2 influenza virus molecule of deactivation is coupled on enzyme plate, show the correct space structure of this albumen, utilize display technique of bacteriophage screening immune nano antibody gene storehouse (camel heavy chain antibody phage display gene pool) with the antigen of this form, thereby obtain the specific nano antibody gene of H3N2, again this gene is gone in intestinal bacteria, can be in the nano antibody strain of E. coli thereby set up.Most importantly, two nano antibodies that we obtain can be for different epitopes, and have good specificity.Obtained nano antibody is carried out to external source transformation, set up a kind of method for quick, for solid basis has been established in the research and development of avian influenza virus diagnostic kit.
Brief description of the drawings
Fig. 1 is the insertion rate detected result in the single domain antibody library of structure, and from left to right the DNA band of gel pore respectively: first is DNA molecular marker, and all the other ducts are the PCR product that detects Insert Fragment, and PCR product band is about 500bp; After testing, the insertion rate in this library reaches more than 90%.
Fig. 2 is after ni-sepharose purification, the electrophorogram of the SDS-PAGE of two kinds of H3N2 nano antibodies that obtain, and the albumen of its expression is the nucleotide sequence shown in corresponding SEQ ID NO:17 or 18 respectively.Result demonstration, H3N2 nano antibody is through this purge process, more than its purity to 95%.
Fig. 3 is the mode chart that H3N2 nano antibody external source is added vitamin H.
Fig. 4 is the H3N2 nano antibody purifying figure of coupling vitamin H, and what this nano antibody was corresponding is the nucleotide sequence shown in SEQ ID NO:18, and SDS-PAGE runs cementing fruit and shows that its purity is more than 90%.
Fig. 5 is by the nucleotide sequence coded H3N2 nano antibody coupling horseradish peroxidase (HRP) shown in SEQ ID NO:17, as detecting antibody, detects figure for the ELISA to H3N2 antigen.
Fig. 6 is the reaction pattern figures of two strain H3N2 nano antibodies for detection of antigen.
Fig. 7 is double antibodies sandwich method based on the Streptavidin MagneSphere detected result to H3N2 antigen.
Embodiment
The present invention's application is coupled on enzyme plate for H3N2, the correct space structure of display protein matter, with the antigen selection immune nano antibody gene storehouse (camel heavy chain antibody phage display gene pool) of this form, can be in the nano antibody strain of E. coli and obtained.
A first aspect of the present invention, a kind of VHH chain of H3N2 nano antibody is provided, has comprised framework region FR and complementary determining region CDR, described framework region FR is made up of four framework regions, be respectively FRl, FR2, FR3 and FR4, their aminoacid sequence is selected from following aminoacid sequence:
FR1 is the aminoacid sequence shown in SEQ ID NO.1, and FR2 is the aminoacid sequence shown in SEQ ID NO.2, and FR3 is the aminoacid sequence shown in SEQ ID NO.3, and FR4 is the aminoacid sequence shown in SEQ ID NO.4;
Or FR1 is the aminoacid sequence shown in SEQ ID NO.5, FR2 is the aminoacid sequence shown in SEQ ID NO.6, and FR3 is the aminoacid sequence shown in SEQ ID NO.7, and FR4 is the aminoacid sequence shown in SEQ ID NO.8;
Described complementary determining region CDR is made up of three complementary determining regions, CDRl, CDR2 and CDR3 respectively, and their aminoacid sequence is selected from following aminoacid sequence:
CDR1 is the aminoacid sequence shown in SEQ ID NO.9, and CDR2 is the aminoacid sequence shown in SEQ ID NO.10, and CDR3 is the aminoacid sequence shown in SEQ ID NO.11;
Or CDR1 is the aminoacid sequence shown in SEQ ID NO.12, CDR2 is the aminoacid sequence shown in SEQ ID NO.13, and CDR3 is the aminoacid sequence shown in SEQ ID NO.14.
Preferably, described specificity is for the nano antibody of A type H3N2 influenza virus, and it has the aminoacid sequence shown in SEQ ID NO.15 or 16.
Second aspect present invention, a kind of H3N2 nano antibody, it is the nano antibody of specificity for A type H3N2 influenza virus epi-position, comprises two VHH chains with aminoacid sequence shown in SEQ ID NO.15 or 16.
Third aspect present invention, a kind of DNA molecular, its coding is selected from the protein of lower group: the VHH chain of the H3N2 nano antibody described in claim 1 or 2, or H3N2 nano antibody claimed in claim 3.
Preferably, described DNA molecular, it has the DNA sequence dna that is selected from lower group: the aminoacid sequence shown in SEQ ID NO.17 or 18.
A fourth aspect of the present invention, a kind of expression vector, it is containing the nucleotide sequence shown in SEQ ID NO.17 or 18.
A fifth aspect of the present invention, a kind of host cell, it can express the nano antibody for A type H3N2 influenza virus.
H3N2 nano antibody external source is added vitamin H and coupling horseradish peroxidase, and is applied to the research and development of quick detection kit.
Below in conjunction with specific embodiment, further set forth the present invention.
The structure in embodiment 1:H3N2 nano antibody library:
(1) the A type H3N2 influenza natural antigen of 0.1mg deactivation is mixed with freund's adjuvant equal-volume, an Xinjiang two-humped camel of immunity, weekly, immunity 7 times altogether, the nano antibody of stimulation B cell expressing antigen-specific; After (2) 7 immunity finish, extract 100 mL camel peripheral blood lymphocytes and extract total RNA; (3) synthesize cDNA and utilize nest-type PRC amplification VHH; (4) utilize restriction enzyme PstI and NotI enzyme cut 20 ug pComb3 Vector for Phage Display (Biovector supply) and 10 ug VHH and connect two fragments; (5) connection product is converted into electricity and turns in competent cell TG1, build H3N2 nano antibody library and measure storage capacity, storage capacity size is 1.0 × 10
9.Meanwhile, 24 clones of our random pickings carry out bacterium colony PCR detection, and result shows that the insertion rate in built library is more than 90%, and Fig. 1 shows bacterium colony PCR result.
Embodiment 2: the nano antibody screening process for H3N2:
(1) will be dissolved in 100 mM NaHCO
3, 20 ug deactivation H3N2 antigens in pH 8.2 are coupled on NUNC enzyme plate, 4 DEG C of placements are spent the night; Within (2) second days, add 100 uL 0.1% caseins, room temperature is sealed 2 h; After (3) 2 h, add 100 uL phages (5 × 10
11tfu immunity camel nano antibody phagocytosis is shown gene pool), room temperature effect 1 h; (4) wash 5 times with 0.05% PBS+Tween-20, to wash non-specific phage off; (5) with 100 mM TEA(triethylamine) will dissociate down with the phage of H3N2 specific binding, and the e. coli tg1 cell of infection in logarithmic phase growth, cultivate 1 h for 37 DEG C, generation purifying phage are for the screening of next round, identical screening process repeats 3-4 wheel, progressively obtains enrichment.
Embodiment 3: the single positive colony of enzyme-linked immunoassay method (ELISA) screening specificity with phage:
(1) in the Tissue Culture Dish that contains phage from the screening of above-mentioned 3-4 wheel, the TB substratum of selecting 96 single bacterium colonies and being inoculated in the penbritin that contains every milliliter of 100 microgram (contains 2.3 grams of potassium primary phosphates in 1 liter of TB substratum, 12.52 gram dipotassium hydrogen phosphate, 12 grams of peptones, 24 grams of yeast extracts, 4 milliliters of glycerine) in, grow to after logarithmic phase, add the IPTG of final concentration 1 mmole, 28 DEG C of overnight incubation.(2) utilize osmose process to obtain and slightly carry antibody, and antibody is transferred in antigen coated elisa plate, at room temperature place 1 hour.(3) wash away unconjugated antibody with PBST, add the anti-mouse-anti HA of mouse anti-HA tag antibody(antibody, purchased from Beijing CoWin Bioscience Co., Ltd.), at room temperature place 1 hour.(4) wash away unconjugated antibody with PBST, add anti-mouse alkaline phosphatase conjugate(goat anti-mouse alkali phosphatase enzyme mark antibody, purchased from the prompt Science and Technology Ltd. of Amy), at room temperature place 1 hour.(5) wash away unconjugated antibody with PBST, add alkaline phosphatase nitrite ion, on ELISA instrument, at 405nm wavelength, read absorption value.(6) when sample well OD value is greater than control wells OD value when more than 3 times, be judged to positive colony hole.(7) bacterium in positive colony hole is turned and shake in the LB liquid that contains every milliliter of 100 microgram to extract plasmid and check order.
Analyze the gene order of each clone strain according to sequence alignment software Vector NTI, by CDR1, CDR2, the strain that CDR3 sequence is identical is considered as same clone strain, and the different strain of its sequence is considered as different clone strains, finally has the different antibody of 6 strains.The aminoacid sequence of the VHH chain of its antibody is respectively as shown in SEQ ID NO:37,38,39,40,41 or 42.
Embodiment 4: nano antibody is at Host Strains expression in escherichia coli, purifying:
(1) the plasmid electricity of different clone strains that sequencing analysis obtains is above transformed in intestinal bacteria WK6, and is coated on the culture plate that LA+glucose contain penbritin and glucose 37 DEG C of overnight incubation; (2) select in the LB nutrient solution that single colony inoculation contains penbritin at 5 mL 37 DEG C of shaking table overnight incubation; (3) spending the night in bacterial classification to 330 mL TB nutrient solution of inoculation 1 mL, 37 DEG C of shaking tables are cultivated, and cultivate while reaching 0.6-0.9 to OD value, add IPTG, 28 DEG C of shaking table overnight incubation; (4) centrifugal, receive bacterium; (5) utilize osmose process, obtain antibody crude extract; (6) can prepare purity through nickel post ion affinity chromatography and reach more than 90% albumen.
Embodiment 5: nano antibody external source is added vitamin H:
(1) use
ncoi and
bstEtwo kinds of restriction enzymes of II, by the plasmid enzyme restriction of two clone strains that obtain, are subcloned into VHH fragment on pBAD carrier; (2) by the carrier successfully constructing and pBiRA plasmid corotation WK6 competent cell, after electricity turns, cell is cultivated to 1h at 37 DEG C of shaking tables, get on the plate culture medium that 200ul is coated on LA+glucose 37 DEG C of overnight incubation; (3) select in the LB nutrient solution that single colony inoculation contains penbritin at 5 mL 37 DEG C of shaking table overnight incubation; (4) spending the night in bacterial classification to 330 mL TB nutrient solution of inoculation 1 mL, 37 DEG C of shaking tables are cultivated, and cultivate while reaching 0.6-0.9 to OD value, add D-Biotin, and making its final concentration is 5mM, and adds IPTG, 28 DEG C of shaking table overnight incubation; (5) centrifugal, receive bacterium; (6) utilize osmose process, obtain antibody crude extract; (7) through Streptavidin
Mutein Matrix purifying, can prepare purity and reach more than 90% albumen.
Embodiment 6: nano antibody coupling horseradish peroxidase (HRP):
(1) take 5mg HRP and be dissolved in 1mL distilled water, add the NaIO of freshly prepared 0.5mL 0.1M/L
4, place 15-30min for 4 DEG C; (2) add 2.5% ethylene glycol 1mL, room temperature is placed 30-60min; (3) add nano antibody 1mg to be marked, and regulate pH to 9.0,4 DEG C of placements are spent the night; (4) add sodium borohydride (5mg/mL) 0.02mL, mix and be placed on 4 DEG C of 3h; (5) saturated ammonium sulphate is removed free HRP, then changes liquid with 1xPBS ultrafiltration, and the centrifugal 30min of 3000rpm, removes throw out, and supernatant is enzyme labelled antibody.(6) whether ELISA detection enzyme labelled antibody can be used.
Embodiment 7: the Streptavidin MagneSphere coupling single domain antibody double antibodies sandwich method detectable antigens based on vitamin H:
(1) double antibodies sandwich method detects two strain nano antibodies and whether identifies different epi-positions, the different epi-positions that two strain H3N2 nano antibodies shown in the present can specific recognition H3N2; (2) get 120ul Streptavidin MagneSphere (each sample 20ul magnetic bead), be placed on magnetic frame, remove protection liquid, then with 1xPBS washing 5 times, add nano antibody (2ug/mL) the room temperature reaction 30min of 1mL coupling vitamin H; (3) smooth, on magnetic frame, suck raffinate, with PBST washing 10 times, then with 0.1%BSA washing 2 times, add 1mL 0.1%BSA sealing 2h; (4) on magnetic frame, suck raffinate, with PBST washing 10 times, add the antigen of different concns: 0ng/mL, 10ng/mL, 50ng/mL, 100ng/mL, 500ng/mL, 1000ng/mL, every kind of concentration antigen volume is 500 μ L, room temperature reaction 1h; (5), with PBST washing 15 times, add the nano antibody (1ng/ μ L) of 500ul coupling HRP, room temperature reaction 30min; (6) PBST washing 25 times, distilled water washing 2 times, each sample adds 100 μ L nitrite ions, and reaction 3-5min, adds 50 μ L 2M H
2sO
4termination reaction, is placed in microplate reader and surveys A450
nmlight absorption value.
The above is only the preferred embodiment of the present invention; be noted that for those skilled in the art; under the premise without departing from the principles of the invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
The present invention utilizes the A type H3N2 influenza virus natural antigen immunity camel of deactivation, obtains high-quality immune nano library of antibody genes.Then the A type H3N2 influenza virus molecule of deactivation is coupled on enzyme plate, show the correct space structure of this albumen, utilize display technique of bacteriophage screening immune nano antibody gene storehouse (camel heavy chain antibody phage display gene pool) with the antigen of this form, thereby obtain the specific nano antibody gene of H3N2, again this gene is gone in intestinal bacteria, can be in the nano antibody strain of E. coli thereby set up.Most importantly, two nano antibodies that we obtain can be for different epitopes, and have good specificity.Obtained nano antibody is carried out to external source transformation, set up a kind of method for quick, for solid basis has been established in the research and development of avian influenza virus diagnostic kit.
SEQUENCE LISTING
<110> Nantong Egens Biotechnology Co., Ltd.
<120> specificity is for the nano antibody of A type H3N2 influenza virus and the application in diagnosis thereof
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ctg caa ttg aac agc ctg aaa act gag gac acg gcc atg tat atc tgt 288
Leu Gln Leu Asn Ser Leu Lys Thr Glu Asp Thr Ala Met Tyr Ile Cys 85 90 95
gca aaa ggg gat tat cgt ggt ggt tac tac tac ccg caa acg tca cag 336
Ala Lys Gly Asp Tyr Arg Gly Gly Tyr Tyr Tyr Pro Gln Thr Ser Gln 100 105 110
ttc agg ggc cag ggg acc cag gtc acc gtc tcc tca 372
Phe Arg Gly Gln Gly Thr Gln Val Thr Val Ser Ser
115 120
Claims (8)
1. a species specificity is for the nano antibody of A type H3N2 influenza virus, comprise framework region FR and complementary determining region CDR, it is characterized in that, described framework region FR is made up of four framework regions, be respectively FRl, FR2, FR3 and FR4, their aminoacid sequence is selected from following aminoacid sequence:
FR1 is the aminoacid sequence shown in SEQ ID NO.1, and FR2 is the aminoacid sequence shown in SEQ ID NO.2, and FR3 is the aminoacid sequence shown in SEQ ID NO.3, and FR4 is the aminoacid sequence shown in SEQ ID NO.4;
Or FR1 is the aminoacid sequence shown in SEQ ID NO.5, FR2 is the aminoacid sequence shown in SEQ ID NO.6, and FR3 is the aminoacid sequence shown in SEQ ID NO.7, and FR4 is the aminoacid sequence shown in SEQ ID NO.8;
Described complementary determining region CDR is made up of three complementary determining regions, CDRl, CDR2 and CDR3 respectively, and their aminoacid sequence is selected from following aminoacid sequence:
CDR1 is the aminoacid sequence shown in SEQ ID NO.9, and CDR2 is the aminoacid sequence shown in SEQ ID NO.10, and CDR3 is the aminoacid sequence shown in SEQ ID NO.11;
Or CDR1 is the aminoacid sequence shown in SEQ ID NO.12, CDR2 is the aminoacid sequence shown in SEQ ID NO.13, and CDR3 is the aminoacid sequence shown in SEQ ID NO.14.
2. specificity according to claim 1, for the nano antibody of A type H3N2 influenza virus, is characterized in that, it has the aminoacid sequence shown in SEQ ID NO.15 or 16.
3. a H3N2 nano antibody, is characterized in that, it is the nano antibody of specificity for A type H3N2 influenza virus epi-position, comprises two VHH chains with aminoacid sequence shown in SEQ ID NO.15 or 16.
4. a DNA molecular, is characterized in that, its coding is selected from the protein of lower group: the VHH chain of the H3N2 nano antibody described in claim 1 or 2, or H3N2 nano antibody claimed in claim 3.
5. DNA molecular according to claim 4, is characterized in that, it has the DNA sequence dna that is selected from lower group: the aminoacid sequence shown in SEQ ID NO.17 or 18.
6. an expression vector, is characterized in that, it is containing the nucleotide sequence shown in SEQ ID NO.17 or 18.
7. a host cell, is characterized in that, it can express the nano antibody for A type H3N2 influenza virus.
8. H3N2 nano antibody claimed in claim 3, is characterized in that, described H3N2 nano antibody external source is added vitamin H and coupling horseradish peroxidase.
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Cited By (3)
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| CN105367655A (en) * | 2015-12-11 | 2016-03-02 | 广东工业大学 | Hand-foot-and-mouth disease resistant specific nanobody and titer determination method thereof |
| CN106188283A (en) * | 2015-05-07 | 2016-12-07 | 北京科卫临床诊断试剂有限公司 | The nano antibody of type A avian influenza H7N2 and application thereof |
| CN114907475A (en) * | 2022-05-31 | 2022-08-16 | 中国科学院合肥物质科学研究院 | Nano antibody for resisting influenza A virus M2 antigen and application thereof |
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| WO2013030604A1 (en) * | 2011-09-02 | 2013-03-07 | Health Protection Agency | Influenza virus antibody compositions |
| CN103755804A (en) * | 2014-01-27 | 2014-04-30 | 南通市伊士生物技术有限责任公司 | Nano antibody for type A H3N2 influenza virus and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106188283A (en) * | 2015-05-07 | 2016-12-07 | 北京科卫临床诊断试剂有限公司 | The nano antibody of type A avian influenza H7N2 and application thereof |
| CN106188283B (en) * | 2015-05-07 | 2020-03-17 | 北京科卫临床诊断试剂有限公司 | Nano antibody of avian influenza A H7N2 and application thereof |
| CN105367655A (en) * | 2015-12-11 | 2016-03-02 | 广东工业大学 | Hand-foot-and-mouth disease resistant specific nanobody and titer determination method thereof |
| CN114907475A (en) * | 2022-05-31 | 2022-08-16 | 中国科学院合肥物质科学研究院 | Nano antibody for resisting influenza A virus M2 antigen and application thereof |
| CN114907475B (en) * | 2022-05-31 | 2023-06-02 | 合肥中科长木生物科技有限公司 | Nanometer antibody for resisting influenza A virus M2 antigen and application thereof |
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