CN109021101A - Anti- PAX8 protein monoclonal antibody and application thereof - Google Patents

Anti- PAX8 protein monoclonal antibody and application thereof Download PDF

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Publication number
CN109021101A
CN109021101A CN201811015866.4A CN201811015866A CN109021101A CN 109021101 A CN109021101 A CN 109021101A CN 201811015866 A CN201811015866 A CN 201811015866A CN 109021101 A CN109021101 A CN 109021101A
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monoclonal antibody
pax8
oti6h8
application
kit
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Inventor
何为无
马东晖
魏海涛
叶露
张乾坤
扈晓敏
王成林
任琪
钮倩
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WUXI ORIGENE BIO-TECHNOLOGY CO LTD
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WUXI ORIGENE BIO-TECHNOLOGY CO LTD
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Publication of CN109021101A publication Critical patent/CN109021101A/en
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57449—Specifically defined cancers of ovaries
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Organic Chemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present invention relates to field of biotechnology, disclose a kind of hybridoma cell strain (deposit number is CGMCC No.15591), and the monoclonal antibody OTI6H8 that thus hybridoma cell strain generates.The invention further relates to monoclonal antibody OTI6H8 to prepare the application in the immune detection tool for detecting PAX8 albumen, application of the immunohistochemical kit and monoclonal antibody OTI6H8 of the OTI6H8 containing monoclonal antibody in the kit that preparation is used for marked tumor.Monoclonal antibody of the present invention can in conjunction with PAX8 protein-specific, and with other intracellular albumen no cross reactions, significantly improve specificity, the accuracy and reliability of the detection of PAX8 protein immunization.

Description

Anti- PAX8 protein monoclonal antibody and application thereof
Technical field
The present invention relates to oncology and medical domain, and in particular to can specific bond PAX8 albumen monoclonal antibody The method and purposes of OTI6H8, the cell strain for generating the monoclonal antibody and the application antibody for tumour Identification of The Origin.
Background technique
Matching box gene -8 (Paired box gene 8, PAX8) is thyroid gland, urogenital tract, kidney and other organs Important transcription regulaton factor in growth course, the constitutive expression in thyroid gland, the tissue such as kidney, in the upper of normal ovarian Chrotoplast is not expressed.But it is positive in core dye in endometroid carcinoma of ovary and clear cell carcinoma, and in mucosal oophoroma, mammary gland And it is hardly expressed in other non-gynecological primary tumors.These expression characteristics of PAX8 are that base has been established in its clinical application Plinth.The expression situation of albumen in immunohistochemistry (IHC) Pathological experiment detection tumour cell is clinically commonly used at present, however The core of IHC experiment is the monoclonal antibody of binding proteins specific, and the superiority and inferiority of performance directly decides the spirit entirely detected Sensitivity and specificity.Therefore, a kind of higher monoclonal antibody for PAX8 albumen of binding specificity is developed to detect IHC PAX8 expression has great importance.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of monoclonals of the higher PAX8 albumen of binding specificity to resist Body, and its preparing the application in the immune detection tool for detecting PAX8 albumen.
The present invention provides a kind of hybridoma cell strains, and it is commonly micro- to be preserved in China Committee for Culture Collection of Microorganisms Bio-Centers (referred to as CGMCC), the deposit date is on April 26th, 2018, deposit number was CGMCC No.15591.
It is thin by above-mentioned hybridoma the present invention also provides a kind of monoclonal antibody OTI6H8 for specifically binding PAX8 albumen Born of the same parents' strain generates.
Monoclonal antibody of the present invention the preparation method is as follows:
(1) building of recombinant expression carrier: according to PAX8ORF nucleotide sequence (PAX8ORF nucleotide sequence such as SEQ ID Shown in NO.1,714bp；PAX8 amino acid sequence is as shown in SEQ ID NO.2)
Design primer PCR amplification PAX8ORF 898bp to 1350bp bit sequence, gene two sides introduce limitation respectively Property restriction enzyme site SgfI and MluI, be inserted into expression vector pET23-N-His, building PAX8 amino acid sequence the 300th to the 450 recombinant expression plasmid pET23a-rPAX8；Upstream amplification primer sequence, SEQ ID NO.3:CACGCGATCGCG GATCCTCACTCACCCTTC downstream amplification primer sequence SEQ ID NO.4:ACCGACGCGT CTA CAGATGGTCAAAGGCCG
(2) expression and purification of PAX8 recombinant protein: by PAX8 recombinant expression plasmid Transformed E .Coli cell, cracking centrifugation Supernatant is obtained, affinity chromatography column purification obtains the PAX8 recombinant protein of purifying；
(3) screening and preparation of monoclonal antibody: Balb/c mouse is immunized using the PAX8 recombinant protein of above-mentioned purifying, is taken Mouse spleen cells are merged with SP2/0 cell, and limiting dilution assay obtains monoclonal, and it is thin that ELISA method screens positive hybridoma Born of the same parents obtain the hybridoma cell strain that can secrete anti-PAX8 specific antibody, are named as OTI6H8, subtype identification IgG2b；Pass through Serum free medium prepares antibody, obtains PAX8 monoclonal antibody OTI6H8 by affinity chromatography column purification.Pass through respectively Western Blot, immunohistochemical experiment verify the sensitivity and specificity of the monoclonal antibody.
The present invention also provides monoclonal antibody OTI6H8 in preparing the immune detection tool for detecting PAX8 albumen Application.
Specifically, the immune detection tool is kit, chip or test paper.
In the particular embodiment, the present invention provides a kind of immunologic combined detection reagent kit, including above-mentioned monoclonal are anti- Body OTI6H8 can detect the expression situation of PAX8 in histocyte.
The present invention also provides application of the said monoclonal antibody in the kit that preparation is used for marked tumor.Wherein institute State the tumour being proliferated and the expression of PAX8 is closely related that tumour specifically refers to tumour cell, including but not limited to oophoroma group It knits.
Compared with prior art, the present invention provides a kind of hybridoma cell strain (deposit number CGMCCNo.15591), And the monoclonal antibody OTI6H8 that thus hybridoma cell strain generates.The present invention also provides monoclonal antibody OTI6H8 to make The application being ready for use in the immune detection tool of detection PAX8 albumen, the immunohistochemical kit of the OTI6H8 containing monoclonal antibody, And application of the monoclonal antibody OTI6H8 in the kit that preparation is used for marked tumor.Monoclonal antibody of the present invention can It is true to reflect PAX8 protein expression level in tumour cell in conjunction with PAX8 protein-specific, it can be applied to distinguish ovary endometrial Sample cancer and clear cell carcinoma.
Preservation information
The classification naming of hybridoma cell strain OTI6H8 for preservation are as follows: the hybridoma of PAX8 mouse monoclonal antibody Strain；
Depositary institution's full name: China Committee for Culture Collection of Microorganisms's common micro-organisms center；
Depositary institution's abbreviation: CGMCC；
Depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica；
Preservation date: on April 26th, 2018；
Deposit number: CGMCC No.15591.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technical description to be briefly described.
Fig. 1 shows the design of 1 cloning site of embodiment as schemed, and wherein shading part is the area ORF；
Fig. 2-5 show 4 formalin of embodiment fix, the normal kidney of paraffin embedding, kidney, normal thyroid, thyroid gland The histogenic immunities group such as tumour result figure (primary antibody is PAX8 monoclonal antibody OTI6H8)；
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described, Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all Belong to the scope of protection of the invention.
The building of embodiment 1, PAX8 recombinant expression plasmid
It is with the plasmid RC200651 (containing PAX8ORF1350bp) obtained from Biotechnology Co., Ltd of Aureal Dongyuan County of the U.S. Template designs two primers and introduces restriction enzyme site SgfI and MluI respectively, is cloned into expression vector pET23a-N-His, establishes PAX8 recombinant expression plasmid.Cloning site design is as shown in Figure 1.
The expression and purification of embodiment 2, PAX8 recombinant protein
1, Transformed E .coli cell: being added Plasmid DNA and gently mix after 100ul competent cell is placed on ice to melt, ice 42 DEG C of heat shock 90s after bath 30min, are then continued ice bath 1-2min.The fresh nonreactive LB culture of 500ul is added in super-clean bench Base takes appropriate bacterium solution to be spread evenly across on antibiotic plate, culture dish is inverted in 37 after 37 DEG C of shaking tables are incubated for 45min Overnight incubation in DEG C constant incubator.
2, lytic cell: picking monoclonal is in fresh culture, and 37 DEG C, 200rpm cultivates to OD value and reach 0.4~0.6 When be added IPTG (final concentration 1mM) Fiber differentiation 7h.Thalline were collected by centrifugation, and thallus then is resuspended with lysis buffer, and ultrasound is broken 12000rpm is centrifuged 20min at 4 DEG C after broken 20min, collects supernatant.A small amount of supernatant protein is taken to make WB mirror with anti-His antibody It is fixed.
3, affinity chromatography column purification: nickel column is balanced with buffer, simultaneously by loading after 0.45 μm of membrane filtration of supernatant Outflow is collected, is received respectively with the unbonded albumen of buffer elution removal finally with the elution of the imidazoles containing various concentration SDS-PAGE identification is carried out after collection, satisfactory elution albumen is merged, 10% glycerol is added, recombination PAX8 egg after purification It is white to be identified with SDS-PAGE.
The preparation and screening of embodiment 3, PAX8 monoclonal antibody
The PAX8 protein fragments of the purifying generated according to standard method recombination are used to (tie up tonneau in Beijing to Balb/c mouse Magnificent experimental animal Technology Co., Ltd.) it is immunized.The specific method is as follows:
1, animal immune: purified PAX8 antigen is emulsified with complete Freund's adjuvant, using subcutaneous or intraperitoneal injection side 6-8 week old Balb/c mouse is immunized in method, and immunizing dose is 50 μ g/, and progress is immune for the second time after two weeks at interval, with not exclusively not Family name's adjuvant emulsion, immunizing dose are 50 μ g/.It is immune that tail blood is taken to measure serum titer with ELISA method gradient dilution afterwards twice；Root Booster immunization is determined whether according to result, is chosen the highest mouse of antibody titer and is carried out cell fusion.
2, cell fusion: myeloma cell uses the sp2/0 in the source Balb/c, and logarithmic growth phase is in when fusion；It takes Immune mouse spleen, is made lymphocyte single cell suspension；Mouse spleen lymphocyte is mixed with myeloma cell with 1:5-1:10, 37 DEG C 50%PEG (PH 8.0) 1mL is added dropwise, incomplete culture medium and remaining terminate liquid is added, HAT is added after abandoning supernatant in centrifugation Culture medium, which suspends, to be mixed, and MC constant volume to 50mL is dispensed into 3.5cm culture dish, is put in wet box, is placed in 37 DEG C, 5%CO2It is permanent It is cultivated in warm incubator.
3, screen and clone: fusion selected cell clone in 7-10 days, carried out ELISA survey using PAX8 purification of recombinant proteins Examination.Mark cell strain number.Limiting dilution, 5-6 days measurement ELISA values after each limiting dilution, picking are carried out to positive hole cell The OD280 positive is worth higher monoclonal hole and carries out limiting dilution, until ELISA measures 96 orifice plates, hardened fruit is the positive entirely.Picking The positive is worth high monoclonal singling.It is OTI6H8 that it, which corresponds to fusion plate cell strain,.
4, the preparation and purification of ascites monoclonal antibodies: the male Balb/c mouse peritoneal of 10-12 week old injects 0.5ml norphytane, Every mouse is injected intraperitoneally with 1mL syringe and wash the monoclonal cell suspension being resuspended through PBS after a week, cell dosage for 5 × 106/ only, every strain antibody makes a call to 2 mouse.Ascites, centrifuging and taking supernatant are collected after mouse ascites accumulation, affinity chromatography carries out abdomen Water purifying selects corresponding column material according to antibody subtype, and the monoclonal antibody that cell strain OTI6H8 is generated is IgG2b, using protein G into Row purifying.Monoclonal antibody concentration mensuration after purification, WB are detected, dispense, are frozen at -20 DEG C.
Embodiment 4, the immunohistochemistry that monoclonal antibody OTI6H8 is primary antibody detect
(1), experimental method:
1, the people's lymph node tissue and human tonsil's tissue block for taking formalin fixed carry out paraffin embedding, use Finesse histotome is sliced, and tissue thickness is 6 μm.
2, dewaxing and aquation: analyzing pure 3 × 10min of dimethylbenzene, dehydrated alcohol 3 × 10min, 95% ethyl alcohol 5min, and 85% Ethyl alcohol 5min, 75% ethyl alcohol 5min, deionized water impregnate 3min × 3 time
3, antigen retrieval buffers (0.01M, pH6.0 sodium citrate buffer) pressure cooker hot high pressure is added and repairs 3min, to height When pressure pot temperature is down to about 90 DEG C, pressure cooker is opened, takes out sample, then cooled to room temperature.Deionized water impregnates 3min × 3 times.
4, using 3% hydrogen peroxide deactivation tissue endogenous peroxydase, it is stored at room temperature 10min.Deionized water is impregnated 5min × 3 time.
5, confining liquid (+5% Normal Goat Serum of PBS+5% skimmed milk power) is added, 37 DEG C of incubation 60min.
6, confining liquid is removed, is not rinsed, is added PAX8 monoclonal antibody (OTI6H8), thinner ratio: 1:150 is carried out using confining liquid Dilution.It is placed in wet box, 37 DEG C of incubation 60min.PBST (0.1%Tween-20) is washed 2 times, washs 5min every time.PBST (0.02%Tween-20) is washed 1 time, washs 5min every time.
7,1,37 DEG C of reagent of Polink- kit 2 (Catlog No.D37-15) incubation 10-20 minutes is added dropwise.Use PBS Washing 3 times, each 5min.2,37 DEG C of reagent of Polink-2 kit (Catlog No.D37-15) incubation 10-20 minutes is added dropwise, It is washed 3 times using PBS, each 5min.
8, it develops the color using DAB solution (Zhong Shan Golden Bridge ZLI-9019), develop the color 3~10min.Distill water washing.
9, haematoxylin redyeing nucleus 2min, distilled water rinsing, the differentiation of 1% hydrochloric acid.Distilled water rinses 3 times, is stored at room temperature 1min。
10, it is dehydrated and transparent: 75% ethyl alcohol 5min, 100% ethyl alcohol 5min x 3 times, 85% ethyl alcohol 5min, 95% ethyl alcohol 5min, 100% 3 × 5min of ethyl alcohol；3 × 5min of dimethylbenzene, neutral gum mounting.
11, microscopy is shown in Fig. 2-5.
(2), experimental result:
By the result of Fig. 2~5 as it can be seen that there is the nuclear staining of specific region in normal kidney, thyroid gland and tumor tissues.Knot Fruit and PAX8 positioning in the cell and tissue expression specificity are consistent, show that monoclonal antibody OTI6H8 can be used for immuning tissue The level of chemical detection PAX8 albumen.
<110>Wuxi Origene Bio-tech Co., Ltd.
<120>anti-PAX8 protein monoclonal antibody and application thereof
<210> 1
<211>2961
<212> DNA
<213>artificial sequence
<400> 1
ORIGIN
1 ATGCCTCACA ACTCCATCAG ATCTGGCCAT GGAGGGCTGA ACCAGCTGGG AGGGGCCTTT
61 GTGAATGGCA GACCTCTGCC GGAAGTGGTC CGCCAGCGCA TCGTAGACCT GGCCCACCAG
121 GGTGTAAGGC CCTGCGACAT CTCTCGCCAG CTCCGCGTCA GCCATGGCTG CGTCAGCAAG
181 ATCCTTGGCA GGTACTACGA GACTGGCAGC ATCCGGCCTG GAGTGATAGG GGGCTCCAAG
241 CCCAAGGTGG CCACCCCCAA GGTGGTGGAG AAGATTGGGG ACTACAAACG CCAGAACCCT
301 ACCATGTTTG CCTGGGAGAT CCGAGACCGG CTCCTGGCTG AGGGCGTCTG TGACAATGAC
361 ACTGTGCCCA GTGTCAGCTC CATTAATAGA ATCATCCGGA CCAAAGTGCA GCAACCATTC
421 AACCTCCCTA TGGACAGCTG CGTGGCCACC AAGTCCCTGA GTCCCGGACA CACGCTGATC
481 CCCAGCTCAG CTGTAACTCC CCCGGAGTCA CCCCAGTCGG ATTCCCTGGG CTCCACCTAC
541 TCCATCAATG GGCTCCTGGG CATCGCTCAG CCTGGCAGCG ACAAGAGGAA AATGGATGAC
601 AGTGATCAGG ATAGCTGCCG ACTAAGCATT GACTCACAGA GCAGCAGCAG CGGACCCCGA
661 AAGCACCTTC GCACGGATGC CTTCAGCCAG CACCACCTCG AGCCGCTCGA GTGCCCATTT
721 GAGCGGCAGC ACTACCCAGA GGCCTATGCC TCCCCCAGCC ACACCAAAGG CGAGCAGGGC
781 CTCTACCCGC TGCCCTTGCT CAACAGCACC CTGGACGACG GGAAGGCCAC CCTGACCCCT
841 TCCAACACGC CACTGGGGCG CAACCTCTCG ACTCACCAGA CCTACCCCGT GGTGGCAGAT
901 CCTCACTCAC CCTTCGCCAT AAAGCAGGAA ACCCCCGAGG TGTCCAGTTC TAGCTCCACC
961 CCTTCCTCTT TATCTAGCTC CGCCTTTTTG GATCTGCAGC AAGTCGGCTC CGGGGTCCCG
1021 CCCTTCAATG CCTTTCCCCA TGCTGCCTCC GTGTACGGGC AGTTCACGGG CCAGGCCCTC
1081 CTCTCAGGGC GAGAGATGGT GGGGCCCACG CTGCCCGGAT ACCCACCCCA CATCCCCACC
1141 AGCGGACAGG GCAGCTATGC CTCCTCTGCC ATCGCAGGCA TGGTGGCAGG AAGTGAATAC
1201 TCTGGCAATG CCTATGGCCA CACCCCCTAC TCCTCCTACA GCGAGGCCTG GCGCTTCCCC
1261 AACTCCAGCT TGCTGAGTTC CCCATATTAT TACAGTTCCA CATCAAGGCC GAGTGCACCG
1321 CCCACCACTG CCACGGCCTT TGACCATCTG
//
<210> 2
<211>986
<212> PRT
<213>artificial sequence
<400> 2
ORIGIN
1 MPHNSIRSGH GGLNQLGGAF VNGRPLPEVV RQRIVDLAHQ GVRPCDISRQ LRVSHGCVSK
61 ILGRYYETGS IRPGVIGGSK PKVATPKVVE KIGDYKRQNP TMFAWEIRDR LLAEGVCDND
121 TVPSVSSINR IIRTKVQQPF NLPMDSCVAT KSLSPGHTLI PSSAVTPPES PQSDSLGSTY
181 SINGLLGIAQ PGSDKRKMDD SDQDSCRLSI DSQSSSSGPR KHLRTDAFSQ HHLEPLECPF
241 ERQHYPEAYA SPSHTKGEQG LYPLPLLNST LDDGKATLTP SNTPLGRNLS THQTYPVVAD
301 PHSPFAIKQE TPEVSSSSST PSSLSSSAFL DLQQVGSGVP PFNAFPHAAS VYGQFTGQAL
361 LSGREMVGPT LPGYPPHIPT SGQGSYASSA IAGMVAGSEY SGNAYGHTPY SSYSEAWRFP
421 NSSLLSSPYY YSSTSRPSAP PTTATAFDHL

Claims (6)

1. a kind of monoclonal antibody, which is characterized in that it is special with pairing box gene -8 (Paired box gene 8, PAX8) The opposite sex combines；
The monoclonal antibody is generated by hybridoma cell strain OTI6H8, and deposit number is CGMCC No.15591.
2. monoclonal antibody as described in claim 1 is in preparation for the immune of dividing tissue lymphatic endothelial and blood vessel endothelium Application in detection instrument.
3. application according to claim 2, the immune detection tool is reagent, kit, chip or test paper.
4. a kind of immunologic combined detection reagent kit, which is characterized in that including monoclonal antibody described in claim 1.
5. application of the monoclonal antibody as described in claim 1 in the kit that preparation is used for tagged tissue cell.
6. application according to claim 5, the histocyte is oophoroma, human thyroid carcinoma.
CN201811015866.4A 2018-09-01 2018-09-01 Anti- PAX8 protein monoclonal antibody and application thereof Pending CN109021101A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110577595A (en) * 2019-08-09 2019-12-17 无锡傲锐东源生物科技有限公司 anti-TTF 1 protein monoclonal antibody and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160370370A1 (en) * 2011-05-10 2016-12-22 Biocare Medical, Llc Systems and Methods for Anti-PAX8 Antibodies

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160370370A1 (en) * 2011-05-10 2016-12-22 Biocare Medical, Llc Systems and Methods for Anti-PAX8 Antibodies

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PETRA I.LORENZO ET AL.: "Immunohistochemical assessment of Pax8 expression during pancreatic islet development and in human neuroendocrine tumors", 《HISTOCHEMISTRY AND CELL BIOLOGY》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110577595A (en) * 2019-08-09 2019-12-17 无锡傲锐东源生物科技有限公司 anti-TTF 1 protein monoclonal antibody and application thereof

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