CN105695418B - A kind of monoclonal antibody and application of structure specific nuclease FEN 1 - Google Patents
A kind of monoclonal antibody and application of structure specific nuclease FEN 1 Download PDFInfo
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- CN105695418B CN105695418B CN201610149086.3A CN201610149086A CN105695418B CN 105695418 B CN105695418 B CN 105695418B CN 201610149086 A CN201610149086 A CN 201610149086A CN 105695418 B CN105695418 B CN 105695418B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
-
- 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
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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- 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
- G01N33/57411—Specifically defined cancers of cervix
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- 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
- G01N33/57415—Specifically defined cancers of breast
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- 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
- G01N33/57423—Specifically defined cancers of lung
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- 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
- G01N33/57434—Specifically defined cancers of prostate
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- 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
- G01N33/57446—Specifically defined cancers of stomach or intestine
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- 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
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- Chemical Kinetics & Catalysis (AREA)
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- Genetics & Genomics (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of monoclonal antibody of structure specific nuclease FEN 1 and application, the hybridoma cell strain for secreting the monoclonal antibody is additionally provided.The monoclonal antibody can be used for detecting the kit of cancer cell, and the cancer cell is preferably prostate cancer, breast cancer, stomach cancer cell, neuroblastoma, cancer of pancreas or lung cancer.
Description
Technical field
The present invention relates to the monoclonal antibody of structure specific nuclease (FEN1) a kind of, secrete the miscellaneous of the monoclonal antibody
Hand over the application of tumor cell strain and the antibody.
Background technique
The maintenance of Genome stability and the duplication of DNA and reparation require a series of endonuclease and circumscribed
The participation of enzyme, structure specific nuclease 1 (flap endonuclease 1, FEN1) are exactly one of them.FEN1 can identify spy
Fixed DNA bifurcation structure, and cut off the single-chain nucleic acid containing free 5 ' end.During DNA replication dna, FEN1 is circumscribed by its
Enzyme, endonuclease activity eliminate the last one ribonucleotide of Okazaki fragments front end RNA primer, and in DNA reparation, FEN1 is with it
Inscribe enzyme activity takes part in the repair process (1) of damaged base.Due to importance of the FEN1 in DNA replication dna and reparation, once
Encoding the gene of FEN1, defect will lead to FEN1 the abnormal of content increased or reduced in vivo, or occur functional
Variation, so as to cause the unstable of genome, leads to the generation of tumour or cancer.
FEN1 belongs to a member in XPG/Rad2 nucleic acid family, possesses 5 ' bifurcated restriction endonucleases (FEN), 5 ' excision enzymes
(EXO) and notch relies on the special nucleases of 3 kinds of structures such as endonuclease (GEN).FEN1 gene is in human genome
In be positioned on the 11q12.2 of chromosome (2), contain 2 exons and 1 introne.1141 of its Exon
380 amino acid residues of alkali yl coding FEN1, total molecular weight about 42KD (3).
FEN1 is as a kind of structure specific nuclease, RNA in Okazaki fragments maturation when being primarily involved in DNA replication dna
The excision of primer, long segment base excision repair, the decomposition of DNA secondary structure, the maintenance of DNA replication dna fork and promotion Apoptosis
The processes such as the DNA fragmentation generation of induction.It will form the valvular structure of many similar flaps during these, and FEN1 mainly knows
Not this structure, and cut off by its activity, to maintain the stability of DNA.It, can be with during FEN1 is functioned
Different protein combines, so that influencing it participates in different DNA replication dna and reparation approach.So far, it has been found that at least 20
There is interactions between multiple proteins and FEN1.PCNA is the accessory factors of an archaeal dna polymerase, it is in the cell S phase
Be during DNA replication dna it is required, recombining to the base during the NER of damage dna also has effect.And for FEN1's
Function, PCNA and its combination are particularly significant.28 amino acid regions (AA328-355) of FEN1 have active in conjunction with PCNA
(4).5-50 times of shear efficiency can be improved in FEN1 under conditions of having PCNA to stimulate, after PCNA can increase FEN1 to shearing site
Combination stability, perhaps have stronger shear efficiency (5).FEN1, which is mutated, can change the combination of it and PCNA, reduce it
Activity in duplication will cause DNA replication dna amplification, so as to cause cancer and genetic disease (6).
High expression of the FEN1 on nucleic acid or protein level can have an impact tumour.In metastatic forefront
Adenocarcinoma cell, breast cancer, stomach cancer cell, neuroblastoma, cancer of pancreas, in lung cancer cell line its visible expression up-regulation
(7).By expression of the research FEN1 in prostate cancer, and compare primary prostate cancer, prostatic intraepithelial neoplasia,
Average expression of the expression discovery FEN1 of FEN1 in prostate cancer in benign prostatic hyperplasis and normal prostatic epithelium
Level is 36.7%, is much higher than normal level 13.2%, and is 14.5% in benign prostatic hyperplasis, is swollen in prostatic epithelium
Tumor is 15.4%, this illustrates FEN1 excessively high expression in prostate cancer, and it expresses dedifferenting for the degree and cell increased
It is related.Its result implies that FEN1 may be as a kind of diagnostic markers of prostate cancer.
FEN1 takes part in numerous life processes as a kind of structure specific nuclease, grinds to its structure and function
Study carefully the hot spot of always DNA replication dna and damage research field.For FEN1 and tumour relationship research shows that it is likely to make
The getting up early diagnosis of tumour is used for for a kind of marker of tumour.
Summary of the invention
One aspect of the present invention provides a kind of hybridoma cell line, which is characterized in that the preservation of the hybridoma cell line
Number be CGMCC No.12013.
Another aspect of the present invention provides a kind of structure specific nuclease 1 secreted by hybridoma cell line above-mentioned
Monoclonal antibody, which is characterized in that it being capable of specifically integrated structure specific nucleic acid enzyme FEN1.
It is special in detection target structure that further aspect of the present invention provides hybridoma above-mentioned and monoclonal antibody above-mentioned
Application in specific nuclease 1.
Further aspect of the present invention provides the reagent comprising hybridoma above-mentioned and/or monoclonal antibody above-mentioned
Box.
Further aspect of the present invention provides hybridoma above-mentioned and monoclonal antibody above-mentioned in detection cancer cell
Application in kit, the cancer cell be preferably prostate cancer, breast cancer, stomach cancer cell, neuroblastoma, cancer of pancreas,
Lung cancer, cervical carcinoma.
Detailed description of the invention
Fig. 1, FEN-1 antibody sodium dodecyl sulfate polyacrylamide gel electrophoresis method (SDS-PAGE) detection.
Fig. 2, immune-blotting method difference lysate (CHO-K1, Raji, Raw264.7, C6, PC-12, COS7,3T3,
Hela, Jurkat) in FEN-1 albumen expression (antibody extension rate 1:1000).
The immunofluorescence of Fig. 3, FEN-1 antibody tests (cell line used: HeLa, antibody thinner ratio 1:400).
Specific embodiment
Method therefor is normal applying method unless otherwise instructed in following embodiments.
Reagent, enzyme, carrier and cell strain:
(1) cell strain: HeLa (CCL-2TM)、CHO-K1(CCL-61TM)；Raji(
CCL-86TM)、Raw264.7(TIB-71TM)、C6(CCL-107TM)、PC-12(CRL-
1721TM)、COS7(CRL-1651TM)、 3T3(CRL-1658TM)、Jurkat(TIB-
152TM)。
(2) experimental animal: female BAl BIc/c mouse (Hunan SJA Laboratory Animal Co. , Ltd)；
(3) carrier: pET41a (Novagen)
(4) enzyme and antibody reagent: the various enzymes of carrier construction process and PCR process are purchased from Fermentas, ELISA experiment
Antibody anti-mouse IgG/HRP (Jackson ImmunoResearch) used, ELISA substrate TMB (Sigma), reverse transcription
Kit (Fermentas), BSA are purchased from (Beijing Yuan Heng Golden Horse Biotechnology Co., Ltd)
(5) other reagents are that domestic analysis is pure unless otherwise instructed.
Embodiment 1: the acquisition of hybridoma cell strain FEN1 and its monoclonal antibody of generation
1. prepared by antigen
(1) target gene is obtained
Trizol Reagent reagent extracts HeLa cell total rna, by total serum IgE reverse transcription is cDNA and using cDNA as template
PCR amplification FEN1 gene.
(2) recombinant expression carrier is constructed
It will recycle after the PCR product double digestion of step (1) acquisition, connect under T4DNA connection enzyme effect into expression vector
PET41a, construction recombination plasmid pET41a-FEN1.Carrier is after digestion and sequencing identification for converting expression bacterium.
(3) the expression strain containing recombinant expression plasmid is obtained
Recombinant plasmid transformed Escherichia coli Rosetta (DE3) competent cell that step (2) are obtained, with containing sulfuric acid
The LB solid medium of kanamycins screens, and picking monoclonal is used for antigen presentation.
(4) expression and purity
Rosetta (DE3) bacterial colony containing expression plasmid is inoculated in the LB that 10ml contains kanamycin sulfate
In fluid nutrient medium, 37 degree, 220rpm cultivates 10hr.Bacterium solution is inoculated in the fresh LB of 200ml by 1:100 extension rate to cultivate
Base, culture to OD value are 0.6, and 0.1mM IPTG inducing expression is added, and 20 degree of 5hr collect bacterium solution ultrasonication, pure from supernatant
Change recombinant antigen FEN1.
2. the preparation and purification of monoclonal antibody
(1), animal is immunized
6-8 week old female BAl BIc/c mouse is generally used, carries out inoculation three times according to the immunization method pre-established.
It is immune for the first time that the 0.2ml emulsion (weight that 100ul is purified is subcutaneously injected with every mouse groin of 2ML syringe
Histone FEN1 (dilutes)+100ul Freund's complete adjuvant, antigen containing 100ug with 1 × PBS).
Second immunization interval two weeks, with every mouse peritoneal injection 0.2ml emulsion of 2ML syringe, (100ul was purified
Recombinant protein FEN1 (with 1 × PBS dilute)+100ul incomplete Freund's adjuvant, antigen containing 100ug).
Third time immunization interval two weeks, with every mouse groin subcutaneous injection injection 0.2ml emulsion of 2ML syringe
(the recombinant protein FEN1 of 100ul purifying (dilutes)+100ul incomplete Freund's adjuvant, antigen containing 100ug with 1 × PBS).
Potency is surveyed according to time of fusion arrangement, takes blood examination to survey after third time is 7-10 days immune, with conventional western method
Potency is detected, potency is selected to reach the mouse of 1:1000 for merging.
Booster immunization merges first 3 days, carries out booster immunization to spleen mouse to be taken, mouse peritoneal injection 200ul liquid (contains
60ug antigen, solvent are 1 × PBS, pH7.4).
(2), cell fusion
Mouse is put to death using eyeball excise depletion method, sterile working takes out spleen, is put into the plate of 200 mesh stainless (steel) wire of band
Middle grinding, prepares cell suspension.Ready homology myeloma cell SP2/0 is mixed in a certain ratio with mouse boosting cell
(1:5-1:10), and be added and promote fusion agent polyethylene glycol (50%).Using HAT Selective agar medium, the choosing of hybridoma is carried out
The culture of selecting property and screening.
Detect Hybridoma Cell Culture supernatant with ELISA method: by antigen coating buffer, (pH 9.6,0.05M carbonate are slow
Fliud flushing) it is diluted in 8ug/ml 96 hole elisa Plates of addition, the hole 50ul/ is put into 4 DEG C of refrigerator-freezers and is coated with overnight.Coating buffer is discarded, is used
PBST (phosphate buffer, pH7.4) board-washing three times, is added confining liquid 3%BSA-PBST and (3% bovine serum albumin(BSA) is added
Phosphate buffer), the hole 100ul/, 37 DEG C are incubated for 1 hour.Discard confining liquid, PBST (phosphate buffer, pH7.4) board-washing
Once, plank is dried.ELISA Plate 100ul/well is added in Hybridoma Cell Culture supernatant, while PBST is added (phosphate is slow
Fliud flushing) it is used as negative control.37 DEG C are incubated for 1 hour.Supernatant is abandoned, three times with board-washing machine PBST board-washing, the anti-diluted is added
IgG/HRP, 100ul/well, 37 DEG C of mouse are incubated for 1 hour.It abandons supernatant and three times with board-washing machine PBST board-washing adds substrate
TMB100ul/well, RT are protected from light 37 DEG C, and 50ul terminate liquid (2mol/L sulfuric acid) is added after 10-15min.It is surveyed using microplate reader
It is fixed, microplate reader setting are as follows: 450nm colorimetric.It is compared with negative control, if it is being exactly positive on 2.5 times of OD value.Finally screen
The hybridoma cell strain of the best anti-FEN1 of one plant of potency is obtained, 7H8, subtype identification IgG1 are named as.
The positive hybridoma cell strain 7H8 filtered out is subjected to monoclonal screening (limiting dilution assay), acquisition can generate height
The hybridoma cell clone of potency (WB thinner ratio > 1:5000) monoclonal antibody.Hybridoma cell strain is expanded and is cultivated, and is frozen
Conservation.The positive hybridoma cell is anti-human FEN1 hybridoma cell line 7H8, which has been stored in China Microbiological
Culture presevation administration committee common micro-organisms center, address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number
Are as follows: CGMCC No.12013, preservation date are as follows: on January 15th, 2016.
(3) preparation and purification of monoclonal antibody
The cell strain 7H8 that step (2) obtain is seeded to BALB/c mouse abdominal cavity, prepares ascites, it is then pure from ascites
Change antibody (obtained antibody is named as anti-FEN1).
The purifying of anti-FEN1 monoclonal antibody: Protein G affinity purification is used.It will be good with PB solution equilibria
Protein G filler is fitted into purification column, and the ascites through the diluted monoclonal antibody anti-FEN1 of PB solution is added and crosses column.On
After sample, pillar is washed to flowing through value < 0.01 liquid OD with PB solution.Then it is washed with the glycine-HCI solution of 0.1M pH3.0
It is de-, collect the solution of entire eluting peak.Eluent is concentrated by dialysis and is changed to PBS solution.SDS-PAGE is the results show that pure
Antibody purity is in 95% or more (referring to Fig. 1) after change.
Embodiment 2: Identification of Monoclonal Antibodies and application
1. the immune-blotting method of the anti-FEN1 monoclonal antibody (anti-FEN1) of mouse is identified
The cell pyrolysis liquid of various tumor cell lines is extracted, total protein concentration is adjusted to 2mg/ml, loading to 12%
In SDS-PAGE glue hole, the hole 20ug/, by the protein delivery to pvdf membrane in gel after electrophoresis, after the closing of 5% skimmed milk power,
The anti-FEN1 antibody being prepared with embodiment 1 is incubated for: the anti-that 1 step of embodiment (3) prepare and purify is added
FEN1, working concentration are respectively 1:1000, then the Goat anti-mouse antibodies of HRP label, working concentration is added in 4 degree of overnight incubations
1:20000, in 37 degree of reaction 1hr, there is the band (referring to fig. 2) of 45KD, is consistent with document report in each concentration gradient, card
This bright antibody is the specific antibody of anti-FEN1；Antibody dilution gradient reaches 1:1000, and (antibody working concentration can reach 0.2ug/
Ml) there is clear band, be able to detect FEN1 in CHO-K1 (Lane 1), HeLa (Lane 2), Raji (Lane 3), RAW246.7
(lane 4), C6 (Lane 5), PC-12 (Lane 6), COS7 (Lane 7), 3T3 (Lane 8), Jurkat (Lane 9) cell
In expression.
2. dyeing immunofluorescence cell is identified
Hela cell is washed twice with PBS, and the fixed 30min of 4% paraformaldehyde room temperature, PBS is rinsed, with 0.5%Triton X-
100PBS room temperature permeabilization 20min.Then 5%BSA PBS is added to close nonspecific reaction.1 step of embodiment (3) system is added
Anti-FEN1 monoclonal antibody 1:400,37 degree of incubation 30min standby and purify.It rinses, the mountain sheep anti mouse of FITC label is added
IgG, room temperature, which is protected from light, is incubated for 1hr.The free secondary antibody of removal, PBS are rinsed, fluorescence mountant mounting, and fluorescence microscopy under the microscope, is used
Blue light excitation observation green florescent signal.Obvious green fluorescence is presented into nucleus for experimental result (Fig. 3) clear view, fixed
Position is consistent with document report.Illustrate that the specific antibody of this anti-FEN1 can be used for the expression of Immunofluorescence test observation FEN1 and determine
Position.
1.Shen,B.,Singh,P.,Liu,R.,Qiu,J.,Zheng,L.,Finger,L.D.,Alas,S.,(2005)
Multiple but dissectible functions of FEN-1 nucleases in nucleic acid
processing,genome stability and diseases.Bioessays 27,717-729.
2.Hiroka LR,Harrington JJ,Gerhard DS,Lieber MR,Hsieh CL(1995)Sequence
od human FEN1,a structure-specific endonuclease,and chromosomal localization
of the gene(FEN1)in mouse and human.Genomics25:220-225.
3.Pan MH, Du J, Zhang JY, Huang MH, Li T, Cui HJ, Lu C (2011) Cloning of the
flap endonuclease-1 gene in Bombyx mori and indentification of an
antiapoptotic function.DNA Cell Biol 30:763-770.
4.Gary R, Ludwig DL, Cornelius HL, MacInnes MA, Park MS (1997) The DNA
repair endonuclease XPG binds to proliferating cell nuclear antigen(PCNA)and
shares sequence elements with the PCNA-binding regions of the FEN1 and
cyclin-dependent kinase inhibitor p21.J Biol Chem 272:24522-24529
5.Tom S,Henricksen LA,Bambara RA(2000)Mechnism whereby proliferating
cell nuclear antigen stimulates flap endonuclease 1.J Biol Chem 275:10498-
10505
6.Hosfield DJ, Mol CD, Shen B, Tainer JA (1998) Structure of the DNA
repair and replication endonuclease and exonuclease FEN1:coupling DNA and
PCNA binding to FEN1 activity.Cell 95:135-146
7.Singh P,Yang M,Dai H,Yu D,Huang Q,Tan W,Kernstine K.H.,Lin D,Shen B
(2008)Over expression and hypomethylation of flap endonuclease 1 gene in
breast and other cancers.Mol.Cancer Res.6,1710-1717
Claims (5)
1. a kind of hybridoma cell line, which is characterized in that the deposit number of the hybridoma cell line is CGMCC No.12013.
2. a kind of monoclonal for the structure specific nuclease FEN 1 secreted by hybridoma cell line described in claim 1 resists
Body, which is characterized in that it being capable of specifically integrated structure specific nucleic acid enzyme FEN1.
3. hybridoma cell line described in claim 1 and monoclonal antibody as claimed in claim 2 detect target structure in preparation
Application in the kit of specific nucleic acid enzyme FEN1.
4. a kind of reagent comprising hybridoma cell line described in claim 1 and/or monoclonal antibody as claimed in claim 2
Box.
5. hybridoma cell line described in claim 1 and monoclonal antibody as claimed in claim 2 detect cancer cell in preparation
Kit in application；The cancer cell is prostate cancer, breast cancer, stomach cancer cell, neuroblastoma, cancer of pancreas, lung
Cancer, cervical carcinoma.
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| CN103415771A (en) * | 2011-03-11 | 2013-11-27 | 霍夫曼-拉罗奇有限公司 | Fen1 as marker for chronic obstructive pulmonary disease (copd) |
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Non-Patent Citations (6)
| Title |
|---|
| Identification of flap-structure specific endonuclease 1 as a factor involved in long-term memory formation of aversive learning.;Lorena Saavedra-Rodríguez et al.;《J Neurosci》;20090506;第29卷(第18期);第5726-5737页 * |
| Identification of flap-structure specific endonuclease 1 as a factor involved in long-term memory formation of aversive learning;Lorena Saavedra-Rodríguez et al.;《J Neurosci》;20090506;第29卷(第18期);第5726-5737页 * |
| 周婷等.结构特异性核酸内切酶FEN1及其与肿瘤的关系.《南京师大学报（自然科学版）》.2014,第37卷(第2期),第7-14页. * |
| 王越.雌激素对乳腺癌细胞中FEN1表达的影响及其机制研究.《中国优秀硕士学位论文全文数据库医药卫生科技辑》.2011,(第4期),第8页倒数第1行. * |
| 结构特异性核酸内切酶FEN1及其与肿瘤的关系;周婷等;《南京师大学报（自然科学版）》;20140630;第37卷(第2期);第7-14页 * |
| 雌激素对乳腺癌细胞中FEN1表达的影响及其机制研究;王越;《中国优秀硕士学位论文全文数据库医药卫生科技辑》;20110415(第4期);第8页倒数第1行 * |
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