CN102060912A - Amyloid protein oligomer conformation type epitope polypeptide and application thereof - Google Patents
Amyloid protein oligomer conformation type epitope polypeptide and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of peptide-containing medical preparations, in particular relating to an amyloid protein oligomer conformation type epitope polypeptide and an application thereof. The epitope polypeptide is used for preparing an antibody which can identify a plurality of amyloid protein oligomers for the diagnosis, treatment and experimental research of amyloid protein diseases. The invention provides the amyloid protein oligomer conformation type epitope polypeptide (PEOZ) the amino acid sequence of which is Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys or is obtained by replacing, deleting or adding one or more of amino acid residues to the amino acid sequence. The epitope polypeptide can effectively stimulate an organism to generate the specific antibody which can identify amyloid protein oligomers and the antibody can inhibit the aggregation of the amyloid protein and the cytotoxicity.
Description
Technical field
The invention belongs to the pharmaceutical product field that contains peptide, be specifically related to a kind of amyloid oligomer conformation type antigen epitope polypeptide and application thereof; The present invention is applied to discern diagnosis, treatment and the experimental study of the Antibody Preparation amyloid disease of multiple amyloid oligomer.
Background technology
For many years, the multiple disease that causes of the gathering of amyloid or polypeptide self has caused increasing harm to human beings'health.Nontoxicity of some protein own or toxicity are very little, but they self can be gathered into oligomer (oligomer) with toxic action or fibrous material (fibril) and cause a series of diseases, as the A Zihaimo disease (being commonly called as senile dementia) that causes by Beta-amyloid (A-Beta) (Alzheimer ' disease, AD), the Parkinson's disease that cause by alpha-synuclein (Parkinson ' disease, PD), at least ten surplus kinds of encephalopathics of the humans and animals that comprises mad cow disease that causes by prion protein (Prion protein (PrP)), the at least 9 kinds of heredity nerve degenerative diseases that comprise Huntington Chorea (Huntington ' s disease) that cause by the polypeptide that contains poly glumine (PolyQ), by islet amyloid polypeptide (islet amyloid polypeptide, IAPP, type ii diabetes that amylin) causes and the N,O-Diacetylmuramidase (lysozyme) that is caused by long-time dialysis assemble the disease that deposition causes etc.Wherein, be AD and PD and type ii diabetes to the human health risk maximum.Medical statistics shows, 5～6% suffer from AD among the over-65s the elderly in China and the American-European countries, and sickness rate rises year by year.This disease has been listed in and has caused dead the fourth-largest disease, is only second to heart trouble, cancer and apoplexy.Other have an appointment 1% over-65s the elderly suffers from PD.And the number of suffering from type ii diabetes can reach more than 5% of total population.These diseases have caused increasing harm to human beings'health, and the basic reason of its morbidity (or partly cause) is the gathering of amyloid or polypeptide self.
Studies show that the gathering of different proteins starts from protein (or polypeptide) monomer of false folding or sex change at first, the formation of monomer polypeptide interchain hydrogen bond has caused the polymerization of protein molecular.At first form spherical oligomer by the solubility of electronics or the about 3-10 nm of the observable size of atomic force microscope, some oligomer can be further combined with becoming crooked flexible filament (protofibril), and then to form diameter be the smooth surface of 6-10 nm or fiber in the shape of a spiral.The protein of non-homology finally can form the protein polymer with analog structure.The fiber that is formed by various amyloid aggregations such as A-Beta, alpha-synuclein and amylin all contains " cross-Beta " structure, and wherein the skeleton of Beta lamella formation is vertical with the fiber longitudinal axis, and the hydrogen bond net in the skeleton is parallel with the longitudinal axis.Polypeptide is arranged as parallel Beta chain in the Beta lamella, in the Beta lamella, amino acid all has accurate position.Even the oligomer of different sources also has similar constitutional features, the oligomer that oligomer specific recognition antibody can form with the amyloid monomer by different sources combines, and does not combine with their monomer and fiber.This shows that oligomer can form the distinctive epitope of common oligomer that is independent of outside its amino acid primary sequence by the protein polypeptide skeleton.Colloid gold particle simulation A-Beta 40 oligomers that the Glabe laboratory applications is connected with A-Beta 40 prepared not only can combine with A-Beta 40 and A-Beta 42 oligomers and can with the polyclonal antibody (A11) of the oligomer specific combination of formation such as alpha-synuclein, IAPP, PolyQ, PrP, Insulin, Lysozyme, this antibody can also effectively suppress the cytotoxicity of all these oligomers.
What the past people thought the amyloid disease always is to be caused by the insoluble fiber that protein aggregation becomes.A large amount of in recent years studies show that, the crucial paathogenic factor of many amyloid diseases is the less water-soluble oligomeric things of volume.The disease that is caused by the albumen oligomer exists similar mechanism, and promptly cell membrane damage, oxidative stress, mitochondrial function imbalance, necrocytosis, signal transmission are unusual etc.But people it be unclear that the detailed mechanism that oligomer influences the normal physiological activity of cell.How are the topological framework of oligomer and epitope thereof, and these all are that people demand the problem inquiring into and solve urgently how could to suppress its cytotoxicity etc. effectively.Thereby use the gathering that active and passive immunological technique treatment AD suppress A-Beta, and the removing of quickening A-Beta is one of focus of AD research field.
2000-2002 U.S. scientific research personnel uses the vaccine that A-Beta makes and has carried out an animal experiment and a clinical trial phase.Experimental result is encouraging, and laboratory animal and patient's clinical symptom obviously alleviates, and memory is obviously recovered, and pathological change is also significantly gone down.Simultaneously, use anti-A-Beta antibody the passive immunotherapy of laboratory animal has also been received better curative effect.Yet, unfortunately in phase ii clinical trial,, have 6% patient table to reveal meningitic symptom and pathological change, the patient who has even take place dead though most patient still shows good result.Therefore, the experimental study to A-Beta vaccine and antibody stops immediately.However, from above-mentioned research we as can be seen the A-Beta vaccine should affirm the result of treatment of AD.Studies show that the side effect that is caused by the A-Beta vaccine may be the autoimmune response by the T cell mediated of this vaccine-induced body generation, and the generation of this side effect is caused by Th1 rather than the immune response of Th2 type.In order to overcome the side effect of A-Beta vaccine, when vaccine design, should strengthen bringing out B cell response and the immunoreactive generation of Th2 type, and avoid bringing out t cell immune response and the immunoreactive generation of Th1 type.Vaccine based on the design of A-Beta oligomer epitope peptide can easily be taken into account above-mentioned characteristic, also can not induce body to form the monomeric antibody of anti-A-Beta.Back one characteristic can thoroughly be eliminated the possibility that has side effects between the A-Beta monomer in vaccine and the body, and can not influence the normal physiological function of A-beta.Therefore, the peptide vaccine of developing according to A-Beta oligomer epitope will have the potential using value, and obtain the prerequisite that A-Beta oligomer epitope is this peptide vaccine design and development.So, scientific research and clinical in all need to obtain a kind of amyloid oligomer conformation type antigen epitope polypeptide.
Summary of the invention
The object of the present invention is to provide a kind of amyloid oligomer conformation type antigen epitope polypeptide (being called for short PEOZ), its aminoacid sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys, or by above-mentioned aminoacid sequence through replacement, disappearance or add one or several amino acid, and have the polypeptide that suppresses the amyloid aggregation effect.The present invention can produce the specific antibody of discerning the amyloid oligomer by the effective stimulus body, and this antibody can suppress the gathering and the cytotoxicity of amyloid.
In specific embodiments of the invention, the N end of polypeptide of the present invention is used the PEG2000 modification, and (sequence is: PEG-Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys), (sequence is the deletion L-Ala: Cys-Ser-His-Leu-His-Asn-Asn-Cys), Serine is replaced as L-Ala, and (sequence is: Cys-Ala-His-Ala-Leu-His-Asn-Asn-Cys) or insert L-Ala (sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys), show that after testing the PEOZ polypeptide is through some modification, certain amino acid is through displacement, lack or in polypeptide, still can remain with and A11 bonded ability behind the increase amino acid.
In vitro tests shows that polypeptide of the present invention can combine with A11, has proved that described polypeptide is one of pairing epitope of polyclone A11; Described polypeptide has good antigenic characteristic, but the effective stimulus body produces antibody; Described polypeptide is a common epitope on the multiple amyloid oligomer; Described polypeptide antibody can obviously suppress amyloid A b42, PrP, amylin, a-synuclein, lysozyme gathering; Described polypeptide antibody can obviously suppress the cytotoxicity of Ab42, PrP, amylin, a-synuclein, lysozyme.
The present invention also provides described polypeptide to be applied to amyloid oligomer inhibitor.
The present invention also provides described polypeptide to be applied to prepare specific recognition amyloid oligomer antibody.
The present invention also provides described polypeptide to be applied to preparation treatment amyloid disease vaccine.
The present invention has following beneficial effect compared to existing technology:
1. the present invention can produce the specific antibody of discerning the amyloid oligomer by the effective stimulus body, and this antibody can suppress the gathering and the cytotoxicity of amyloid.
2. B cell response and the immunoreactive generation of Th2 type be can strengthen bringing out based on vaccine of the present invention, and t cell immune response and the immunoreactive generation of Th1 type avoided bringing out; Can not induce body to form the monomeric antibody of anti-A-Beta, can thoroughly eliminate the possibility that has side effects between the A-Beta monomer in vaccine and the body, and can not influence the normal physiological function of A-beta.
3. the present invention has better immunogenicity, but the effective stimulus body produces antibody, thereby can be used as good immunogen in amyloid treatment of diseases or prevention aspect the development of vaccine.
Description of drawings:
Fig. 1. PEOZ and A11 bonded ELISA measure;
Fig. 2. measure through replacing, lack or inserted amino acid whose PEOZ and A11 bonded ELISA;
Fig. 3. the antibody after the PEOZ immunity in the mice serum combines with the Ab42 oligomer;
Fig. 4. PEOZ antibody combines with the oligomer of multiple amyloid;
Fig. 5. ThT fluorescent dyeing determination PEOZ antibody suppresses the gathering of amyloid;
Fig. 6. PEOZ antibody suppresses the cytotoxicity of amyloid;
Embodiment:
Embodiment 1:
One peptide species (being called for short PEOZ), its aminoacid sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys, or by above-mentioned aminoacid sequence through replacement, disappearance or add one or several amino acid, and have the polypeptide that suppresses the amyloid aggregation effect.The present invention can produce the specific antibody of discerning the amyloid oligomer by the effective stimulus body, and this antibody can suppress the gathering and the cytotoxicity of amyloid.
The N end of described polypeptide is used the PEG2000 modification, and (sequence is: PEG-Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys), (sequence is the deletion L-Ala: Cys-Ser-His-Leu-His-Asn-Asn-Cys), Serine is replaced as L-Ala, and (sequence is: Cys-Ala-His-Ala-Leu-His-Asn-Asn-Cys) or insert L-Ala (sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys), show that after testing the PEOZ polypeptide is through some modification, certain amino acid is through displacement, lack or in polypeptide, still can remain with and A11 bonded ability behind the increase amino acid.
In vitro tests shows that described polypeptide can combine with A11, has proved that described polypeptide is one of pairing epitope of polyclone A11; Described polypeptide has good antigenic characteristic, but the effective stimulus body produces antibody; Described polypeptide is a common epitope on the multiple amyloid oligomer; Described polypeptide antibody can obviously suppress amyloid A b42, PrP, amylin, a-synuclein, lysozyme gathering; Described polypeptide antibody can obviously suppress the cytotoxicity of Ab42, PrP, amylin, a-synuclein, lysozyme.
Described polypeptide is applied to the amyloid polygalacturonase inhibitor; Also be applied to prepare specific recognition amyloid oligomer antibody; Also be applied to preparation treatment amyloid disease vaccine.
Described polypeptide can produce the specific antibody of discerning the amyloid oligomer by the effective stimulus body, and this antibody can suppress the gathering and the cytotoxicity of amyloid.B cell response and the immunoreactive generation of Th2 type be can strengthen bringing out based on the vaccine of described polypeptide, and t cell immune response and the immunoreactive generation of Th1 type avoided bringing out; Can not induce body to form the monomeric antibody of anti-A-Beta, can thoroughly eliminate the possibility that has side effects between the A-Beta monomer in vaccine and the body, and can not influence the normal physiological function of A-beta.
The method of the synthetic preparation of PEOZ is:
PEOZ is annular seven peptides, (preparation method is referring to Weng C C by the synthetic according to a conventional method preparation of the biochemical company limited of Shanghai gill, Peter D W. Fmoc Solid Phase Peptide Synthesis:A Practical Approach. Oxford:University Press, 2000. 1 ~ 8; Atherton E, Sheppard R C. Solid Phase Peptide Synthesis:A Practical Approach. Oxford:IRL Press, 1989); PEOZ purity is more than or equal to 95%; PEOZ is stored in-20 ° of C, avoids multigelation.
The preparation method that PEOZ is used for special anti-amyloid oligomer antibody is:
The M13 phage immunity new zealand white rabbit of PEOZ polypeptide will be showed.Per 2 all immunity once, each immunity be an individual phagemid/rabbit with antigen.Immunity is 4 times altogether.After last immune 15 days, gather rabbit anteserum.With the sepharose that is connected with the PEOZ polypeptide rabbit anteserum is carried out affinity chromatography, the antibody of the anti-PEOZ of purifying.The antibody of the oligomer of this specific combination amyloid can be used for experimental study, and amyloid treatment of diseases and clinical diagnosis are also had the potential using value.
All experimental datas of PEOZ are all by at least 3 independently experiment acquisitions, and experimental data is expressed as mean number plus-minus standard deviation.The statistical study of data is to use One-way ANOVA software to carry out.
Below in conjunction with embodiment 2-7, further set forth the character of PEOZ.
Embodiment 2:PEOZ combines with polyclonal antibody A11
With 100 μ l PEOZ(1 mg/ holes) wrap by 96 hole enzyme linked immunological microwell plates (not wrap in contrast) by the hole of PEOZ, put 37 ° of C 2 h, seal 2 h with 37 ° of C of 3% BSA, 200 μ L/ holes.Wash plate 3 times with PBS.Every hole adds 100 μ l A11(1:500), behind incubated at room 1 h, with the PBS washing that contains 0.1% Tween-20 3 times.Behind incubated at room 1 h, with the PBS washing that contains 0.1% Tween-20 3 times.Every hole adds the goat-anti rabbit HRP ELIAS secondary antibody (1:5000) after the 100 μ l dilution, behind incubated at room 1 h, with the PBS washing that contains 0.1% Tween-20 3 times.Every hole adds substrate solution TMB 100 μ L, places 37 ° of C 20 min, and every then hole adds 50 μ l, 1 mmol/L sulfuric acid termination reaction, measures OD value (wavelength 450nm) on enzyme-linked immunosorbent assay instrument.PEOZ shown in Figure 1 can be by the A11 specific recognition.PEOZ is the polypeptide that obtains as the screening substrate with A11, and it has confirmed that with combining of A11 it is one of pairing epitope of polyclonal antibody A11.
Embodiment 3: through displacement, lack or inserted amino acid whose PEOZ and still combine with A11
The N end of PEOZ is used PEG2000 modification (abbreviating PEG-PEOZ as), and (sequence is: PEG-Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys), (abbreviate as: PEOZ-A) (sequence is the deletion L-Ala: Cys-Ser-His-Leu-His-Asn-Asn-Cys), Serine is replaced as L-Ala (abbreviating PEOZ-S/A as), and (sequence is: Cys-Ala-His-Ala-Leu-His-Asn-Asn-Cys) or insert L-Ala (abbreviate as: PEOZ-G) (sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys).In order to detect the situation that combines of improved polypeptide and A11, these polypeptide are combined with A11 to competition with having the histidine-tagged PEOZ that does not transform, then with the situation that combines of the ELISA detection latter with A11.Add and be coated with in the hole of A11 adding the PEOZ that has label that 1 mg do not transform mixture 100 ml with the improved PEOZ of 10 mg, detect with ELISA then and have the PEOZ of label and combining of A11, mensuration OD value the results are shown in Figure 2.Show that the PEOZ polypeptide still can combine A11 with the PEOZ effective competition that unmodified is transformed through some modification, certain amino acid through displacement, disappearance or increase amino acid in polypeptide after, cause the PEOZ that has label obviously to reduce (comparing P＜0.01) with the combination of A11 with independent PEOZ positive control.
Embodiment 4:PEOZ can bring out body and produce antibody
Phage (in contrast) immune mouse (5/group) of using the M13 phage of showing the PEOZ polypeptide and not having polypeptide to show, every each immune phage 10
10Individual.Per 2 all immunity once are total to immunity 4 times.The 4th immunity blood sampling in back 15 days, separation of serum.Use the content that ELISA detects PEOZ antibody in the serum.100 μ l Ab oligomer (1 mg/ hole) bag by 96 hole enzyme linked immunological microwell plates, is put 37 ° of C 2 h, seal 2 h(200 μ L/ holes) with 37 ° of C of 3% BSA.Wash plate 3 times with PBS, add the serum of 2000 times of dilutions, behind incubated at room 1 h, with the PBS washing that contains 0.1% Tween-20 3 times.Every hole adds the sheep anti mouse ELIAS secondary antibody (1:3000) after the 100 μ l dilution, behind incubated at room 1 h, with the PBS washing that contains 0.1% Tween-20 3 times.Every hole adds substrate solution TMB 100 μ L, places 37 ° of C 20min, and every then hole adds 50 μ l, 1 mmol/L sulfuric acid termination reaction, measures OD value (wavelength 450nm) (Fig. 3) on enzyme-linked immunosorbent assay instrument.5 mouse of immune PEOZ shown in Figure 2 can both produce the antibody of anti-PEOZ, show that PEOZ has good antigenic characteristic, but the effective stimulus body produces antibody.
The preparation of the immune antibody of embodiment 5:PEOZ and with the combining of each amyloid oligomer
The M13 phage immunity new zealand white rabbit of PEOZ polypeptide will be showed.Per 2 all immunity once, each antigen 10
12Individual phagemid.Immunity is 4 times altogether.After last immune 15 days, gather rabbit anteserum.With the sepharose that is connected with the PEOZ polypeptide rabbit anteserum is carried out affinity chromatography, the antibody of the anti-PEOZ of purifying.
100 μ l Ab42, PrP, amylin, a-synuclein, lysozyme monomer (mon), oligomer (olig) and fiber (fib) (1 mg/ hole) are wrapped respectively by 96 hole enzyme linked immunological microwell plates, the negative contrast of BSA.Anti-with PEOZ antibody as one, anti-with the goat-anti rabbit LgG of HRP mark as two, and by the operation that experimentizes of above-mentioned ELISA method.Fig. 4 shows, PEOZ antibody can combine with the oligomer of Ab, PrP, amylin, a-synuclein respectively, and do not combine (except the lysozyme) with their monomer and fiber, show that PEOZ is a common epitope on the multiple amyloid oligomer, and the monomer of amyloid and fiber there is not this epitope (except the lysozyme) (Fig. 4).
The gathering of embodiment 6:PEOZ antibody vitro inhibition amyloid
1) Ab42, amylin, the a-synuclein with foreign procurement is dissolved to 1 mg/ml with hexafluoroisopropanol (HFIP), and room temperature ultrasonic is handled 10 min, divides to install in the epidorf pipe, with the HFIP that volatilizees in the vacuum, preserves in-20 ° of C then.Place 20 min with preceding Ab42, amylin, the a-synuclein that HFIP was handled in room temperature, add dimethyl sulfoxide (DMSO) (DMSO) then, making each protein concentration is 5 mg/ml, and the PBS damping fluid with 0.02 M pH 7.4 is diluted to desired concn then.PrP, lysozyme then directly are prepared into desired concn with 0.02 M pH, 7.4 PBS.
2) PEOZ antibody is dissolved in the PBS damping fluid of 0.02 M pH 7.4, join Ab42, PrP, amylin, a-synuclein then, (lysozym is the acetate buffer solution of pH2.5 to lysozym solution, all the other samples are the PBS damping fluid of pH7.4) in, make each proteic final concentration be respectively 10,200,20,40,200 μ M.The mol ratio of each albumen and PEOZ antibody is respectively 1:1,10:1,1:1,2:1 and 10:1.With the protein solution that do not add PEOZ antibody as contrast.Ab42, PrP, amylin, a-synuclein, lysozym sample are positioned over 65 ° of C in 37 ° of C(lysozym) placed respectively 1 day, 5 days, 5 hours, 4 days and 6 days.
3) to make its concentration be 5 mM to the phosphate buffered saline buffer that thioflavine (ThT) is dissolved in pH 6.5,50 mM.Sample 20 ml that get after hatching join in the black enzyme plate that contains 180 ml ThT solution.On multi-functional microplate reader, measure the fluorescence intensity of ThT behind the mixing with the emission wavelength of 450 nm excitation wavelengths and 482 nm.The fluorescence intensity of each sample is deducted the background fluorescence of ThT itself and the significant difference situation between the analytic sample.Add the albumen fluorescence intensity that reduces behind the PEOZ antibody and be the inhibiting rate of antibody protein aggregation divided by the albumen fluorescence intensity that does not add antibody.Fig. 5 is the gathering inhibiting rate of PEOZ antibody to Ab42, amylin, a-synuclein, lysozyme, PrP, shows that PEOZ can obviously suppress the gathering of each amyloid.
Embodiment 7:PEOZ antibody suppresses the cytotoxicity of amyloid
(MEM) is made into the individual cells suspension with the SH-SY5Y cell with the substratum that contains 10% foetal calf serum, with 10000 cell inoculations in every hole to 96 porocyte culture plates, every pore volume 100 μ L.Cell was cultivated 24 hours in 37 ° of C, and incubator CO2 concentration is 5%.Every hole adds the sample (mixture of Ab42, PrP, amylin, a-synuclein, lysozyme and PEOZ antibody, amyloid contrast and PBS contrast), Ab42, PrP, amylin, a-synuclein, the proteic final concentration of lysozyme are to be respectively 2,200,5,2,20 μ M, and the final concentration of PEOZ antibody is 1,20,2.5,1,10 μ M.Cell continued to cultivate after 48 hours, and every hole adds MTT solution (5 mg/mL) 10 μ L, and 37 ° of C were hatched 3 hours, stop cultivating, every hole adds 100 μ L dissolution of crystals liquid (10%SDS and 5% isopropylcarbinol are dissolved among the 0.01 M HCL), and 37 ° of C overnight incubation are fully dissolved crystallisate.On multi-functional enzyme-linked immunosorbent assay instrument, measure each hole absorbance value with the 570nm wavelength.With the absorbancy of sample divided by the absorbancy of the cell that does not add sample as the cell activity index, and the significant difference situation between the analytic sample (compare with each amyloid contrast,, P＜0.05; #, P＜0.01), the result shows that PEOZ antibody can obviously suppress the cytotoxicity (Fig. 6) of Ab42, PrP, amylin, a-synuclein, lysozyme.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle 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.
Claims (8)
1. following (a) or polypeptide (b),
(a) its aminoacid sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys,
(b) by the aminoacid sequence in (a) through replacement, disappearance or add one or several amino acid, and have suppress the amyloid aggregation effect by (a) polypeptides derived.
2. polypeptide according to claim 1 is characterized in that, it is modified with PEG2000 for the end of the polypeptide N shown in (a).
3. polypeptide according to claim 1 is characterized in that its aminoacid sequence is: Cys-Ser-His-Leu-His-Asn-Asn-Cys.
4. polypeptide according to claim 1 is characterized in that its aminoacid sequence is: Cys-Ala-His-Ala-Leu-His-Asn-Asn-Cys.
5. polypeptide according to claim 1 is characterized in that its aminoacid sequence is: Cys-Ser-His-Ala-Leu-His-Asn-Asn-Cys.
6. be applied to amyloid oligomer inhibitor according to each described polypeptide of claim 1-5.
7. be applied to prepare specific recognition amyloid oligomer antibody according to each described polypeptide of claim 1-5.
8. be applied to preparation treatment amyloid disease vaccine according to each described polypeptide of claim 1-5.
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| CN102504016A (en) * | 2011-12-08 | 2012-06-20 | 清华大学 | Amyloid fibrillar oligomer conformational epitope polypeptide and application thereof |
| CN102516357A (en) * | 2011-12-02 | 2012-06-27 | 清华大学 | Polypeptide combined with A-beta amyloid and application thereof |
| CN103772487A (en) * | 2012-10-24 | 2014-05-07 | 国家纳米科学中心 | Polypeptide for inhibiting aggregation and toxicity of human amylin, reagent and applications |
| CN110240632A (en) * | 2019-04-18 | 2019-09-17 | 华东理工大学 | A kind of Amylin is affine polypeptide and its application |
| WO2023159970A1 (en) * | 2022-02-28 | 2023-08-31 | 安域生物科技杭州有限公司 | ANTIGEN POLYPEPTIDE MODIFIED ON BASIS OF β-AMYLOID AND USE THEREOF |
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| CN102516357A (en) * | 2011-12-02 | 2012-06-27 | 清华大学 | Polypeptide combined with A-beta amyloid and application thereof |
| CN102516357B (en) * | 2011-12-02 | 2014-06-04 | 清华大学 | Polypeptide combined with A-beta amyloid and application thereof |
| CN102504016A (en) * | 2011-12-08 | 2012-06-20 | 清华大学 | Amyloid fibrillar oligomer conformational epitope polypeptide and application thereof |
| CN103772487A (en) * | 2012-10-24 | 2014-05-07 | 国家纳米科学中心 | Polypeptide for inhibiting aggregation and toxicity of human amylin, reagent and applications |
| CN110240632A (en) * | 2019-04-18 | 2019-09-17 | 华东理工大学 | A kind of Amylin is affine polypeptide and its application |
| CN110240632B (en) * | 2019-04-18 | 2022-11-25 | 华东理工大学 | Amylin affinity polypeptide and application thereof |
| WO2023159970A1 (en) * | 2022-02-28 | 2023-08-31 | 安域生物科技杭州有限公司 | ANTIGEN POLYPEPTIDE MODIFIED ON BASIS OF β-AMYLOID AND USE THEREOF |
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