CN101985475A - Novel polyclonal antibody of secretive peptide INM02 and preparation method and use thereof - Google Patents
Novel polyclonal antibody of secretive peptide INM02 and preparation method and use thereof Download PDFInfo
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- CN101985475A CN101985475A CN2009102636311A CN200910263631A CN101985475A CN 101985475 A CN101985475 A CN 101985475A CN 2009102636311 A CN2009102636311 A CN 2009102636311A CN 200910263631 A CN200910263631 A CN 200910263631A CN 101985475 A CN101985475 A CN 101985475A
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- inm02
- polyclonal antibody
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Abstract
The invention belongs to the field of biotechnologies, and provides a novel polyclonal antibody of a secretive peptide INM02 and a preparation method thereof, the polyclonal antibody is prepared by the following steps: (1) emulsifying an INM02 protein with a Freunds Adjuvant Complete, injecting the emulsified material into the back of a white rabbit for primary immunization; (2) reinforcing immunity in the fourth, the seventh, the ninth and the eleventh weeks after the primary immunization; (3) letting blood after the last immunization, and separating an immune serum; (4) adding NaN3 in the immune serum for cryopreservation. The INM02 polyclonal antibody prepared by the method is good in specificity, high in potency, and convenient and quick for detecting distribution and content of the INM02 in protein samples. Researches show that the INM02 has a close relationship with insulin secretion, so that, the invention provides novel ways and methods for treating diabetes mellitus in the future.
Description
Technical field
The invention belongs to biological technical field, be specifically related to polyclonal antibody of novel secretion peptide INM02 and its production and application.
Background technology
2000, on institute of NAS newspaper, first report the gene expression profile of normal people's hypothalmus-pituitary-adrenal axis tissue (Hu RM, et al.Proc Natl Acad Sci USA.2000Aug 15; 97 (17): 9543-8.), set up gene expression profile (Wang XC, et al.Endocr-Relat Cancer, 2004 of insulin human's tumor tissue afterwards again; 11 (2): 295-303.).From insulin human's tumor tissue, be cloned into a new gene, with its called after INM02.
Human INM02 gene chromosomal localization is in 19q13.3-4, and size is 16.5kb, has 12 exons to be separated by 11 introns, and INM02 transcript size is that encoding sequence comprises 762bp about 3000bp, the albumen of 254 amino-acid residues of coding.SignalP software prediction INM02 albumen has a tangible signal peptide at aminoterminal, and ProtFun 2.2 software functions prediction INM02 may belong to the somatomedin class.Northern blotting technology has been set up the distribution expression pattern of INM02 gene, shows that INM02 gene organization expression ratio is more extensive, expresses particularly abundant in pancreas islet, bladder, testis, lungs tissue.This indication INM02 gene is a gene with extremely important function.Yet want definite its function of understanding, it is very essential preparing its polyclonal antibody.
Summary of the invention
The purpose of this invention is to provide polyclonal antibody of novel secretion peptide INM02 and preparation method thereof.The gene clone, genetic expression, protein purification, polyclonal antibody preparation, the INM02 protein ELISA detection method that are specifically related to secrete peptide INM02 are set up.
Another object of the present invention provides the application of above-mentioned polyclonal antibody.
The invention provides the preparation method of polyclonal antibody of novel secretion peptide INM02, this method comprises the steps:
(1), is injected in the white rabbit back and carries out initial immunity with INM02 albumen and Freund's complete adjuvant emulsification;
(2) behind the initial immunity at the 4th, 7,9,11 all booster immunizations;
(3) bloodletting after the last immunity, separating immune serum;
(4) immune serum adds NaN
3Freezing preservation.
Preparation method of polyclonal antibody of the present invention, the INM02 Argine Monohydrochloride sequence in the step (1) is shown in SEQ ID NO 1.
Preparation method of polyclonal antibody of the present invention, the INM02 protein content in the step (1) can be 80-120 μ g, 90 μ g for example, 100 μ g, 115 μ g, etc.
INM02 albumen is dispersed the 20-40 site injection in the preparation method of polyclonal antibody of the present invention, step (1).Preferably, can adopt the 24-36 site injection.
Preparation method of polyclonal antibody of the present invention, in the step (2), booster immunization be with the INM02 albumen of initial immunity equivalent and Freund's incomplete adjuvant emulsification after inject white rabbit.
Preparation method of polyclonal antibody of the present invention behind the separating immune serum, utilizes albumin A affinity chromatography purifying INM02 polyclonal antibody from immune serum.
Immune serum adds NaN in the preparation method of polyclonal antibody of the present invention, step (4)
3Freezing preservation to the concentration 0.01-0.1%.For example, concentration is 0.02%, 0.03%, 0.05%, 0.06%, 0.08%, or the like.Freezing preservation can be adopted-70 ℃ ,-80 ℃ even liquid nitrogen preservation, etc.
The present invention also provides the application of above-mentioned polyclonal antibody, is about to gained INM02 polyclonal antibody and is used to prepare the used antibody of ELISA detection method.
For example, gained INM02 polyclonal antibody is used for determine the proteic location of INM02.Perhaps, gained INM02 polyclonal antibody is used to detect the proteic content of INM02.
The INM02 albumen that is used for immunity among the present invention can adopt genetically engineered or artificial synthesis to obtain, and also can be to contain the proteic gel band of pure INM02 etc.
Use artificial synthesis to obtain INM02 albumen, promptly corresponding amino-acid residue is connected one by one and get final product according to the sequence (the NCBI accession number is AA023975.2) of SEQ ID NO 1.
Using gene engineering and protein engineering are carried out protein expression and purifying, comprise the protein of expressing the INM02 encoding sequence of removing signal peptide, the clone comprises the encoding sequence of the INM02 that removes signal peptide in suitable expression vector, with this carrier transfection host cell, cultivate this host cell under the condition of this dna segment being suitable for expressing, and from culture, reclaim and the required INM02 albumen of purifying.
The preparation of INM02 gene comprises after PCR reaction and carries out purifying among the present invention, and whether nucleotide sequence analysis checking gene is correct.
Dna encoding sequence (see AY194293, wherein CDS is 1027-1791) and expression vector reorganization with the INM02 of removal signal peptide of the present invention form recombinant expression plasmid.The present invention does not limit specific expression plasmid.In a preferred embodiment, the present invention uses prokaryotic expression carrier pPROEX HT.
Simultaneously above-mentioned recombinant prokaryotic expression vector clone is imported suitable prokaryotic host cell according to a conventional method.It the invention is not restricted to specific host cell, as long as can express described recombinant expression vector.In a preferred version, the present invention uses intestinal bacteria ROSSET (DE3) etc.
Expression product of the present invention is present in the cell space of host cell with the form of inclusion body, crack the separation inclusion body, high concentration urea dissolving inclusion body, separation and purification INM02, albumen behind the purifying is carried out the SDS-PAGE electrophoresis, gel is put into after the dyeing of Coomassie brilliant blue dye liquor, the decolouring, cuts off with the aseptic operation blade and contains the proteic purpose band of INM02.
The present invention adopts new zealand white rabbit as immune animal.To contain the proteic gel of INM02 and pulverize, with Freund's complete adjuvant emulsification, multiple injection immune rabbit, immune serum is isolated in the heart blood sampling after the last immunity, adopts Western Blot method to detect the quality and the titre of antibody.
The INM02 polyclonal antibody that utilization of the present invention prepares is used HRP mark INM02 polyclonal antibody simultaneously, as standard substance, has set up the two sandwich ELISA methods that detect INM02 protein content in serum or the cell culture fluid with the INM02 albumen of prokaryotic expression purifying.
The present invention utilizes the INM02 polyclonal antibody and the ELISA method of above-mentioned acquisition, has detected the content of INM02 in human serum and the MIN6 cell culture fluid, finds that INM02 is the novel secretion albumen that can detectedly obtain in serum or MIN6 cell culture fluid.
The INM02 polyclonal antibody that utilization of the present invention prepares adopts the two location of groupization observation INM02 in pancreas in rat of immunity, found that INM02 is positioned in the pancreas islet, at α cell and β cell expression is arranged all.
The present invention utilizes Northern Blot and ELISA method to observe different concns glucose to the INM02 gene of the plain oncocyte MIN6 of mouse islets cell and the influence of protein excretion.Found that the rising along with glucose concn, the INM02 gene and the protein secretion of MIN6 cell increase gradually.
All basic molecular biology operative techniquies are all with reference to " the molecular cloning third edition " in the above technical scheme.
The present invention has used prokaryotic expression carrier-pPROEX HT expression and purification HIS-INM02 fusion rotein has obtained the polyclonal antibody of INM02 behind the immunizing rabbit.Set up the proteic ELISA method of detection INM02, the result is presented in normal people and various disease patient's the serum and has detected the proteic existence of INM02, confirms that INM02 is a kind of novel secretory protein.Preliminary study the relation of INM02 and islet function or diabetes, Northern blotting and immunohistochemistry show that INM02 gene and albumen mainly are expressed in pancreas islet at pancreas, exocrine gland is few.Studies show that of cell levels, glucose can be regulated INM02 expression of gene and proteic secretion, and same low sugar (5.5mM) is compared, and high sugar (25mM) can obviously raise level and the proteic secretion of INM02 of INM02mRNA.The detected result of ELISA shows that the INM02 albumen in the diabetic serum is lower than orthoglycemic people.Above-mentioned INM02 of studies show that and insulin secretion have confidential relation, may be a novel secretion albumen relevant with beta Cell of islet function or diabetes.
Using gene engineering method of the present invention is produced and is obtained good, the efficient and high-quality INM02 polyclonal antibody of specificity, and set up the proteic detection method of INM02 in serum or the cell culture fluid, these lay the foundation for physiological function and the mechanism of action thereof of further inquiring into INM02, for following treatment of diabetes provides new approach.
Embodiment
Expression, purifying and the Polyclonal Antibody Preparation of embodiment 1 INM02
(1) INM02 construction of prokaryotic expression vector
The cDNA and the pET32a that coding are removed the INM02 full-length proteins of signal peptide carry out double digestion (EcoRI and HindIII), connect with the T4 ligase enzyme, set up the INM02 prokaryotic expression carrier.With recombinant vectors pPROEX HT-INM02 transformed into escherichia coli ROSSET (DE3), ampicillin medium screening positive strain, the plasmid enzyme restriction evaluation is extracted in amplification and order-checking confirms that sequence is correct.
(2) expression of INM02
Choose the mono-clonal colony inoculation to 5ml YTA medium, 37 ℃ * 250rpm to OD600=1-2, be seeded to 500ml YTA medium, 37 ℃ * 250rpm to OD600=1-2, add 1M IPTG to final concentration 1mM, 25 ℃ of abduction delivering 3h, 4 ℃ of 4000rpm 20min, collect thalline, the ultrasonic degradation bacterium is also centrifugal, after will going up cleer and peaceful precipitation (0.01%PBS dissolving) and using the mixing of 5 * sample-loading buffer respectively, make the SDS-PAGE electrophoresis, after the Coomassie brilliant blue dyeing, as seen induce in the postprecipitation band that concentrates at the about 32kd of molecular weight place, and expression amount is low in the supernatant.Prompting INM02 protein expression is in inclusion body.
(3) INM02 purifying
To induce the full bacterium in back centrifugal, with BugBuster (Novagen company) and ultrasonic degradation bacterium, recentrifuge, centrifuged deposit dissolves with the 1 * binging buffer that contains 8M urea, and the dissolved supernatant adds through NiSO
4The resin post of handling well, the washing back obtains the good INM02 albumen of purifying with INM02 albumen in 1 * elute buffer column precipitator.
(4) INM02 polyclonal antibody preparation
The INM02 albumen of above-mentioned purifying is carried out the SDS-PAGE electrophoresis, gel is put into the Coomassie brilliant blue dye liquor dyeed about 4 hours, the taking-up gel is put into destainer and was decoloured about 4～8 hours, cuts off required purpose band with the aseptic operation blade, 1 * PBS washing 2 times, 4 ℃ of preservations are standby.Adopt new zealand white rabbit as immune animal.Blood sample collection was to make negative control before white rabbit made immunization.Pulverize containing the proteic gel of 100ug INM02 approximately, with Freund's complete adjuvant emulsification, after hair is scraped at the white rabbit back, disperse the 24-36 site injection, behind the initial immunity 4,7,9,11 weeks respectively with the antigen of equivalent and Freund's incomplete adjuvant emulsification after booster immunization.Heart blood sampling after the last immunity detects antibody titer with Western Blot method.Immune serum is isolated in carotid artery bloodletting in the 10th day after the last immunity, and immune serum adds NaN
3To final concentration 0.02%, be sub-packed in the 1.5ml centrifuge tube ,-80 ° of preservations are standby.Western Blot method detects antibody titers and quality, and the result shows, all meets many anti-preparation requirements.
The purifying of embodiment 2 INM02 polyclonal antibodies
Use albumin A affinity chromatography purifying INM02 polyclonal antibody from rabbit anti-serum.1: 1 by volume ratio adds Binding/Washing Buffer in serum, the Protein A Resin suspension liquid that in the suction attached column, adds 1ml, after treating resin precipitated, drain damping fluid, add 5mlBinding/Washing Buffer and wash resin one time, the serum that in the adsorption column that installs, slowly adds dilution, controlling the post flow is 1ml/min, after sample is crossed post fully, add 30ml Binding/WashingBuffer and wash pillar, in adsorption column, add 10ml Elution Buffer wash-out antibody, collect elutriant, in elutriant, add the Tris-HCl of 1M immediately, adjust pH to 7.4～8.0.
Embodiment 3 HRP mark INM02 polyclonal antibodies
With HRP mark INM02 polyclonal antibody, preparation is used for second antibody of double fastener heart ELISA.Taking by weighing HRP 25mg is dissolved in 1.25% glutaraldehyde solution, in the room temperature standing over night, reacted enzyme solution is crossed Sephadex G-25 chromatography column, use the physiological saline wash-out, flow rate control was collected brown effluent liquid at 1ml/1 minute, be positioned in the 25ml small beaker, slowly stir, get antibody-solutions 5ml to be marked, dropwise add in the enzyme solution under stirring, add 1M PH9.5 carbonic acid buffer 0.25ml, continue to stir 3 hours, and under agitation dropwise added the equal-volume saturated ammonium sulphate, put 4 ℃ 1 hour, 3000rpm centrifugal half an hour, abandon supernatant, throw out is washed secondary with semi-saturation ammonium sulfate, and last throw out is dissolved among the PBS of a small amount of 0.15M PH7.4, above-mentioned solution is packed in the dialysis tubing, to the PBS buffer saline dialysis of 0.15M PH7.4, remove ammonium ion, 10,000rpm removed precipitation in centrifugal 30 minutes, supernatant liquor is enzyme conjugates, after the packing, and stored frozen.
The foundation of the two sandwich ELISA detection methods of embodiment 4 INM02 albumen
Carry out double fastener heart ELISA with the INM02 antibody of above-mentioned albumin A purifying and the antibody of HRP mark, detect the proteic content of INM02 in serum and the cell culture fluid.Carbonate bag with 0.05M PH 9.6 is cushioned liquid with antibody dilution to 20 μ g/ml, in the reacting hole of each polystyrene board, add 0.1ml, 4 ℃ are spent the night, discard solution in the hole, wash each 3 minutes 3 times with the PBS of 0.15M PH7.4, increase serum 0.1ml is in the above-mentioned reacting hole that has wrapped quilt, put 37 ℃ and hatched 1 hour, discard solution in the hole, wash 3 times with the PBS of 0.15M PH7.4, each 3 minutes, add INM02 antibody (1: the 500) 0.1ml of the HRP mark of fresh dilution in each hole, hatched 1 hour, and discarded solution in the hole for 37 ℃, PBS with 0.15MPH7.4 washes 3 times, each 3 minutes, in each reacting hole, add the TMB colour developing liquid 0.1ml of interim preparation, 37 ℃ 30 minutes, in each reacting hole, add 2M sulfuric acid 0.05ml, termination reaction on the ELISA detector, is surveyed each hole OD value in the 450nm place.
Embodiment 5 cellular localizations of INM02 albumen in pancreatic tissue
Preparation rat pancreas tissue paraffin section de, utilize the INM02 polyclonal antibody and the cellular localization of immunohistochemical methods method research INM02 albumen in pancreatic tissue of above-mentioned acquisition, the result shows that INM02 albumen mainly is expressed in pancreas islet at pancreas, α and β cell all have expression, and few at exocrine gland.
Embodiment 6 detects the content of INM02 albumen in serum
Use SignalP 3.0 software predictions and at aminoterminal a tangible signal peptide (MAAASAGATRLLLLLLMAVAAPSRARG, SEQ ID NO 2) is arranged to INM02 albumen.We are in health check-up of Huashan hospital and inpatient, select the low inferior patient of a collection of healthy population and diabetes, hyperthyroidism, carcinoma of the pancreas, lung cancer, bladder cancer, hyperplasia of prostate, prostate cancer, meningioma, schwannoma and gonad function and amounted to 50 people, two sandwich ELISA methods of use setting up have detected the proteic content of INM02 in the above-mentioned serum, with greater than 2 times of values of negative control as positive findings.The result shows that in all detected serum, detected the proteic existence of INM02, that content is the highest is a sarcoma of bladder patient, and that content is minimum is the low patient of gonad function, finds that simultaneously INM02 albumen is also on the low side in diabetic subject's serum.In addition, in the MIN6 cell culture fluid, also detected the proteic existence of INM02.
Embodiment 7 glucose are regulated INM02 expression of gene and proteic secretion
Use low sugar (5.5mM) and high sugar (25mM) to intervene MIN6 cell and the former foster Islet cells 1 hour, 24 hours and 48 hours of being commissioned to train respectively, observe the change situation of INM02 gene and protein excretion.
Northern blotting result shows that compare with low sugar, high sugar can obviously raise INM02 expression of gene in the MIN6 cell, raises 2 times nearly at 1 hour, raises about 3 times at 24 and 48 hours.Simultaneously, ELISA result shows that the proteic secretion of MIN6 cell INM02 that high sugar is cultivated also obviously increases, and compares with low sugar, increases by 2.1 times in 24 hours, increases by 3.2 times in 48 hours.
Real-time PCR result shows, compares with low sugar, and high sugar also can obviously raise INM02 expression of gene in the former foster Islet cells of being commissioned to train, and raises about 4 times at 24 and 48 hours.
Simultaneously, ELISA result shows that the former proteic secretion of foster Islet cells INM02 of being commissioned to train that high sugar is cultivated also obviously increases, and compares with low sugar, increases by 2.3 times in 24 hours, increases by 3.1 times in 48 hours.
This verifies that further INM02 Polyclonal Antibody Preparation of the present invention is successful.
Those skilled in the art can use the present invention to greatest extent.Therefore, above-mentioned preferred specific embodiments should be understood that only to illustrate, but not limits the scope of the invention by any way.
From the above description, those skilled in the art can understand essential characteristic of the present invention at an easy rate.And under the situation that does not depart from its purport and scope, can carry out various changes and improvement to the present invention.
Sequence table
<210>1
<211>254
<212>PRT
<213>Homo?sapiens
<400>1
Met?Ala?Ala?Ala?Ser?Ala?Gly?Ala?Thr?Arg?Leu?Leu?Leu?Leu?Leu?Leu
1 5 10 15
Met?Ala?Val?Ala?Ala?Pro?Ser?Arg?Ala?Arg?Gly?Ser?Gly?Cys?Arg?Ala
20 25 30
Gly?Thr?Gly?Ala?Arg?Gly?Ala?Gly?Ala?Glu?Gly?Arg?Glu?Gly?Glu?Ala
35 40 45
Cys?Gly?Thr?Val?Gly?Leu?Leu?Leu?Glu?His?Ser?Phe?Glu?Ile?Asp?Asp
50 55 60
Ser?Ala?Asn?Phe?Arg?Lys?Arg?Gly?Ser?Leu?Leu?Trp?Asn?Gln?Gln?Asp
65 70 75 80
Gly?Thr?Leu?Ser?Leu?Ser?Gln?Arg?Gln?Leu?Ser?Glu?Glu?Glu?Arg?Gly
85 90 95
Arg?Leu?Arg?Asp?Val?Ala?Ala?Leu?Asn?Gly?Leu?Tyr?Arg?Val?Arg?Ile
100 105 110
Pro?Arg?Arg?Pro?Gly?Ala?Leu?Asp?Gly?Leu?Glu?Ala?Gly?Gly?Tyr?Val
115 120 125
Ser?Ser?Phe?Val?Pro?Ala?Cys?Ser?Leu?Val?Glu?Ser?His?Leu?Ser?Asp
130 135 140
Gln?Leu?Thr?Leu?His?Val?Asp?Val?Ala?Gly?Asn?Val?Val?Gly?Val?Ser
145 150 155 160
Val?Val?Thr?His?Pro?Gly?Gly?Cys?Arg?Gly?His?Glu?Val?Glu?Asp?Val
165 170 175
Asp?Leu?Glu?Leu?Phe?Asn?Thr?Ser?Val?Gln?Leu?Gln?Pro?Pro?Thr?Thr
180 185 190
Ala?Pro?Gly?Pro?Glu?Thr?Ala?Ala?Phe?Ile?Glu?Arg?Leu?Glu?Met?Glu
195 200 205
Gln?Ala?Gln?Lys?Ala?Lys?Asn?Pro?Gln?Glu?Gln?Lys?Ser?Phe?Phe?Ala
210 215 220
Lys?Tyr?Trp?His?Ile?Ile?Leu?Gly?Gly?Ala?Val?Leu?Leu?Thr?Ala?Leu
225 230 235 240
Arg?Pro?Ala?Ala?Pro?Gly?Pro?Ala?Pro?Pro?Pro?Gln?Glu?Ala
245 250
<210>2
<211>27
<212>PRT
<213>Artificial
<400>2
Met?Ala?Ala?Ala?Ser?Ala?Gly?Ala?Thr?Arg?Leu?Leu?Leu?Leu?Leu?Leu
1 5 10 15
Met?Ala?Val?Ala?Ala?Pro?Ser?Arg?Ala?Arg?Gly
20 25
Claims (10)
1. the polyclonal antibody of novel secretion peptide INMO2 is characterized in that containing the aminoacid sequence shown in the SEQ ID NO 1.
2. the Polyclonal Antibody Preparation method of the described novel secretion peptide of claim 1 INMO2 is characterized in that, this method comprises the steps:
(1), is injected in the white rabbit back and carries out initial immunity with INMO2 albumen and Freund's complete adjuvant emulsification;
(2) behind the initial immunity at the 4th, 7,9,11 all booster immunizations;
(3) bloodletting after the last immunity, separating immune serum;
(4) immune serum adds NaN
3Freezing preservation.
3. preparation method of polyclonal antibody as claimed in claim 2 is characterized in that, the INMO2 protein content in the step (1) is 80-120 μ g.
4. preparation method of polyclonal antibody as claimed in claim 2 is characterized in that, INMO2 albumen is dispersed the 20-40 site injection in the step (1).
5. preparation method of polyclonal antibody as claimed in claim 2 is characterized in that, in the step (2), behind the INMO2 albumen and Freund's incomplete adjuvant emulsification of use and initial immunity equivalent, the injection white rabbit carries out booster immunization.
6. preparation method of polyclonal antibody as claimed in claim 2 is characterized in that, behind the separating immune serum, utilizes albumin A affinity chromatography purifying INMO2 polyclonal antibody from immune serum.
7. preparation method of polyclonal antibody as claimed in claim 2 is characterized in that, immune serum adds NaN in the step (4)
3Freezing preservation to the concentration 0.01-0.1%.
8. the described polyclonal antibody of claim 1 is used to prepare the used antibody of ELISA detection method.
9. the described polyclonal antibody of claim 1 is used for determining the proteic location of INMO2.
10. the described polyclonal antibody of claim 1 is used to detect the proteic content of INMO2 in preparation.
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| CN2009102636311A CN101985475A (en) | 2009-07-28 | 2009-12-19 | Novel polyclonal antibody of secretive peptide INM02 and preparation method and use thereof |
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| CN200910055490 | 2009-07-28 | ||
| CN200910055490.4 | 2009-07-28 | ||
| CN2009102636311A CN101985475A (en) | 2009-07-28 | 2009-12-19 | Novel polyclonal antibody of secretive peptide INM02 and preparation method and use thereof |
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| WO2017035176A1 (en) * | 2015-08-24 | 2017-03-02 | The Board Of Trustees Of The University Of Illinois | Method of preventing or treating obesity with an emc10 inhibitor |
| CN108707586A (en) * | 2018-06-19 | 2018-10-26 | 上海伦泽生物科技有限公司 | The monoclonal antibody of anti-reticulon complex subunit 10 and its application |
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| CN108845142A (en) * | 2018-06-19 | 2018-11-20 | 上海伦泽生物科技有限公司 | Application of the EMC10 Protein Detection object in the diagnosis of preparation obesity and scale evaluation and Bariatric effect assessment product |
| CN110331156A (en) * | 2019-05-30 | 2019-10-15 | 深圳大学 | Anti- Mical2 polyclonal antibody and preparation method thereof |
| CN114149498A (en) * | 2020-09-07 | 2022-03-08 | 复旦大学附属华山医院 | Application of monoclonal antibody of anti-human EMC10 in prevention and treatment of type 2 diabetes |
| WO2023010682A1 (en) * | 2021-08-05 | 2023-02-09 | 广东省农业科学院作物研究所 | Antibody capable of detecting nicotiana tabacum agglutinin protein, preparation method therefor and application thereof |
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- 2009-12-19 CN CN2009102636311A patent/CN101985475A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2017035176A1 (en) * | 2015-08-24 | 2017-03-02 | The Board Of Trustees Of The University Of Illinois | Method of preventing or treating obesity with an emc10 inhibitor |
| CN108707586A (en) * | 2018-06-19 | 2018-10-26 | 上海伦泽生物科技有限公司 | The monoclonal antibody of anti-reticulon complex subunit 10 and its application |
| CN108802372A (en) * | 2018-06-19 | 2018-11-13 | 上海伦泽生物科技有限公司 | Detect the kit of reticulon complex subunit 10 in human serum |
| CN108845142A (en) * | 2018-06-19 | 2018-11-20 | 上海伦泽生物科技有限公司 | Application of the EMC10 Protein Detection object in the diagnosis of preparation obesity and scale evaluation and Bariatric effect assessment product |
| CN108802372B (en) * | 2018-06-19 | 2021-05-11 | 上海伦泽生物科技有限公司 | Kit for detecting endoplasmic reticulum protein complex subunit 10 in human serum |
| CN108707586B (en) * | 2018-06-19 | 2022-03-15 | 上海伦泽生物科技有限公司 | Monoclonal antibody of anti-endoplasmic reticulum protein complex subunit 10 and application thereof |
| CN110331156A (en) * | 2019-05-30 | 2019-10-15 | 深圳大学 | Anti- Mical2 polyclonal antibody and preparation method thereof |
| CN110331156B (en) * | 2019-05-30 | 2021-10-12 | 深圳大学 | anti-Mical 2 polyclonal antibody and preparation method thereof |
| CN114149498A (en) * | 2020-09-07 | 2022-03-08 | 复旦大学附属华山医院 | Application of monoclonal antibody of anti-human EMC10 in prevention and treatment of type 2 diabetes |
| CN114149498B (en) * | 2020-09-07 | 2023-12-05 | 复旦大学附属华山医院 | Application of monoclonal antibody against human EMC10 in preventing and treating type 2 diabetes |
| WO2023010682A1 (en) * | 2021-08-05 | 2023-02-09 | 广东省农业科学院作物研究所 | Antibody capable of detecting nicotiana tabacum agglutinin protein, preparation method therefor and application thereof |
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Application publication date: 20110316 |