CN108864272A - A kind of mammaglobin Staphylococal Protein A epitope and its application - Google Patents
A kind of mammaglobin Staphylococal Protein A epitope and its application Download PDFInfo
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Abstract
The invention discloses belong to be related to a kind of mammaglobin Staphylococal Protein A epitope and its application of immunological technique field.The mammaglobin Staphylococal Protein A epitope, amino acid sequence is as shown in sequence table SEQ ID NO.1.Mammaglobin Staphylococal Protein A epitope of the invention is the Mam-A epitope of cross-film, and screens the monoclonal antibody of one plant of target cell membrane Mam-A, identifies above-mentioned epitope.The monoclonal antibody of Mam-A based on cell membrane localization, the monoclonal antibody than existing commercial cell membrane localization Mam-A have higher specificity and affinity.Therefore breast cancer immune detection and targeted therapy based on antibody of the present invention have higher specificity.Since the affinity and specificity of monoclonal antibody of the present invention are significantly higher than commercial antibody, the specificity of breast cancer detection and the targeting of breast cancer treatment and validity can be significantly improved.
Description
Technical field
The invention belongs to immunological technique fields, and in particular to a kind of mammaglobin Staphylococal Protein A epitope and its application.
Background technique
Breast cancer is the most common malignant tumour in women.It is shown according to the related statistical data of International Cancer Research Center, entirely
New breast cancer case accounts for the 22.7% of global female malignant morbidity to ball every year, and it is pernicious swollen that death accounts for all women
The 14.1% of tumor death, the harm of breast cancer has been more than cervical carcinoma, becomes the No.1 tumor disease for threatening women's health.Therefore,
The early diagnosis of breast cancer and the development of new treatment to raising quality of life of patients and extend patient survival meaning weight
Greatly.
In the first-line treatment of breast cancer, the methods of chemotherapy, radiotherapy, surgical operation, targeted therapy are mostly used.However, because
Most of patient with breast cancers are accompanied by lymphatic metastasis, and operation excision cannot eradicate completely, often control with chemicals auxiliary
It treats.Many chemicals confirm there is good anti-tumor capacity in vitro, and ideal antitumor effect is clinically not achieved
Fruit.Classic chemotherapy reaches tumor locus by blood circulation, substantially reduces pharmaceutical activity by intravenous injection administration, drug,
For drug to tumour without specific binding capacity, the drug practical function concentration of tumor locus is lower simultaneously.Preferably to be controlled
Therapeutic effect, it is necessary to improve tumor locus drug concentration by drug usage amount is improved, poisonous side effect of medicine is caused to increase.Cause
This can be improved focal zone administration concentration and reduce toxic side effect using the pharmaceutical preparation with targeting, be before one kind has very much
The novel medicine feeding method of scape, receives more and more attention in recent years.
Mammaglobin A (Mam-A) is a kind of glycoprotein containing 93 amino acid, has mammary gland-specific and in mammary gland
It is generally raised in cancer patient, research reports that its positive rate is up to 70% or more, is acknowledged as potential breast cancer desirable diagnostic
Target.Studies have shown that Mam-A is in early-stage breast cancer diagnosis, transfer and breast cancer in situ and the differentiation of migration breast cancer etc.
Aspect diagnostic is preferable.To the screening studies have shown that Mam-A of breast cancer in situ, positive rate is in early primary breast cancer
70% or more, it is not expressed in the non-breast cancers tissue such as fibroadenoma of breast.The lung's migration and chest migration of breast cancer are marked
The display of this diagnosis research, the positive rate of Mam-A are above vesicular disease fluid protein 15, show that its diagnostic is good.
Mam-A is widely acknowledged as being potential breast cancer treatment molecular target, has important application prospect.However due to it
Distribution of the epitope on cell membrane is unclear so far, and lacks corresponding identification antibody, correlative study secular stagnation not before.
2009, researcher confirmed that a large amount of Mam-A is deposited in the form of cross-film with laboratory facilities bioinformatics technique for the first time
Partial sector is distributed in cell membrane, and can be identified by specific antibody, and reports currently the only one plant of recognizable cell membrane
The commercialization monoclonal antibody mAb304 (MA512638) of upper Mam-A；This be first about Mam-A antigenic determinant on cell membrane
It is distributed and the research as immune guiding drug delivery is reported.2011, Mam-A passed through above-mentioned as target, researcher
Mam-A antibody coupling nir dye is used for the tracing in vivo of breast cancer, and showing Mam-A can be used as targeted delivery of drugs
Molecular target is used for the treatment of breast cancer.In targeted therapy research of the current Mam-A for breast cancer, one plant of tool is only reported
There is the monoclonal antibody of cell membrane recognition capability.Commercialization identifies that cell membrane Mam-A is rare and seriously hinders it in application study from now on.
Summary of the invention
In order to solve the problems, such as that breast cancer clinical diagnosis and targeted therapy lack breast tissue specificity, the present invention provides
A kind of new mammaglobin Staphylococal Protein A epitope, amino acid sequence KELLQEFIDD, as shown in sequence table SEQ ID NO.1.
Above-mentioned mammaglobin Staphylococal Protein A epitope provided by the invention can be applied to prepare the antibody of mammaglobin A.
Further, the antibody is monoclonal antibody.
The present invention also provides the monoclonal antibodies of mammaglobin A a kind of, identify above-mentioned mammaglobin Staphylococal Protein A table
Position.
In order to select the higher epitope of activity, the present invention predicts Mam-A epitope using Biosun biosoftware,
And clonal expression Mam-A overall length antigen and each epitope section antigen.
Mouse is immunized using overall length Mam-A and prepares monoclonal antibody, epitope section antigen is then based on, passes through
The corresponding antigens epitope that ELISA method screening and identification difference monoclonal antibody is identified, and it is thin to breast cancer according to different monoclonal antibodies
The immunofluorescence dyeing feature of born of the same parents identifies different the identified Mam-A albumen of monoclonal antibody in cell on a cellular level
Positioning, obtains above-mentioned epitope, and preliminary screening determines the monoclonal antibody of the Mam-A of target cell membrane.
Mammaglobin Staphylococal Protein A epitope of the invention and its said monoclonal antibody can pass through chemical synthesis or biology
Synthetic method preparation.
The present invention also provides further provide above-mentioned epitope or said monoclonal antibody in preparation prevention or control
The reagent for treating mammaglobin A abnormal expression associated diseases or the purposes in drug.
Further, the present invention also provides above-mentioned epitope or said monoclonal antibody preparation detection, prevention or
Treat the purposes in breast cancer reagent or drug.
In such use, the reagent be immunohistochemistry detection reagent, immunofluorescence detection agent, ELISA detection reagent,
Breast cancer circulating tumor cell captures reagent.
Present invention provides a kind of target compositions for treating breast cancer, include above-mentioned monoclonal antibody.
It further include azithromycin dihydrochloride in above-mentioned composition.
It further include PLGA nano particle in above-mentioned composition, the PLGA nano particle is in conjunction with said monoclonal antibody.
The preparation method of the target composition of above-mentioned treatment breast cancer is：
PLGA nano particle is activated through EDC/NHS processing rear surface carboxyl, the amino with above-mentioned monoclonal antibody
In conjunction with to obtain the nano particle of coupling monoclonal antibody.Doxorubicin hydrochloride is added in PLGA oil phase and is mixed, it is single to prepare coupling
Anti- load medicine particle (mAb-NP).
Beneficial effects of the present invention：
The present invention provides a kind of Mam-A epitopes of cross-film, and screen the Dan Ke of one plant of target cell membrane Mam-A
Grand antibody identifies above-mentioned epitope.The monoclonal antibody of Mam-A based on cell membrane localization, than existing commercial cell
The monoclonal antibody that film positions Mam-A has higher specificity and affinity.Therefore the breast cancer based on antibody of the present invention is exempted from
Epidemic disease detection and targeted therapy have higher specificity.Since the affinity and specificity of monoclonal antibody of the present invention are significantly higher than
Commercial antibody, therefore the specificity of breast cancer detection and the targeting of breast cancer treatment and validity can be significantly improved.
Detailed description of the invention
Fig. 1 is the Mam-A targeting for identifying different epitope monoclonal antibodies.
Fig. 2 is that cell membrane Mam-A targeting monoclonal antibody mAb-6 detects the immunohistochemistry of breast cancer tissue.
Fig. 3 is that the immunolipid magnetic ball of mAb-6 label captures the identified by immunofluorescence of breast cancer cell.
Fig. 4 is the targeting detection that mAb-6 modifies that PLGA carries medicine particle.
Fig. 5 is that Ki67 detects positive cell rate statistical chart.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with the embodiment of the present invention, to this
Technical solution in inventive embodiments is clearly and completely described, it is clear that described embodiment is that a part of the invention is real
Example is applied, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not making creation
Property labour under the premise of every other embodiment obtained, shall fall within the protection scope of the present invention.
Preparation, screening and the characterization of the anti-Mam-A monoclonal antibody of embodiment 1
1, Mam-A Characterization of antigenic epitopes and clonal expression
With the epitope distribution situation of Biosun biosoftware analysis Mam-A, the screening high antigen active of peak value is good
Amino acid sequence segment, epitope is mainly distributed on 4 sections, respectively A section (20-52aa), B section as the result is shown
(43-70aa), C section (56-82aa) and D section (69-93aa).Each section base is inferred according to Escherichia coli optimal codon
Cause synthesizes overall length (1-93aa) and each section antigen gene, obtains corresponding antigens using prokaryotic expression.
2, the preparation, purifying and subtype identification of Mam-A monoclonal antibody
Mam-A overall length antigen adds 50 μ g/ mouse of equivalent Freund's complete adjuvant back and intraperitoneal injection, carries out within the 4th week and the 8th week
2nd, 3 time immune.Myeloma cell is merged with splenocyte according to conventional hybridoma technology, with Mam-A overall length antigen coat
Elisa plate saves clone strain using indirect elisa method screening positive clone.Ascites prepares mouse monoclonal, with Thermo company
Melon Gel Monoclonal IgG Purification Kit carries out antibody purification.Using the source of mouse monoclonal of HBT company
Antibody subtype identification kit carries out antibody subtype identification.14 plants of anti-Mam-A monoclonal antibodies are obtained altogether, and the results are shown in Table 1.
The identification and characterization of the anti-Mam-A monoclonal antibody of table 1.
3, the epitope identification of monoclonal antibody identification
It by above-mentioned four kinds of Mam-A antigen fragments and Mam-A overall length antigen diluent is 5ug/ml with coating buffer, every hole adds
100ul antigen liquid, after 37 DEG C of incubation 1h 4 DEG C overnight, patted dry after washing.Every hole adds confining liquid 100ul, and room temperature closes 1h, washing
After pat dry.Each monoclonal antibody is with 1：1000 ratios are diluted with antibody diluent.The elisa plate being coated with is taken, every hole adds 100ul to dilute
Good antibody, 37 DEG C of incubation 1h are patted dry after washing.The sheep anti mouse secondary antibody of HRP label is with 1：1000 dilutions, every hole add 100ul, and 37
DEG C be incubated for 1h, patted dry after washing.Every hole adds 100ul developing solution, and room temperature is protected from light 15 minutes.Every hole adds 100ul colour developing to terminate
Liquid, microplate reader 450nm measure OD value.
As a result as shown in Table 1,14 plants of monoclonal antibodies identify that epitope difference can be divided into five classes according to it：Monoclonal antibody mAb-1,
2,3,4 A section, epitope 20-52aa are only identified；MAb-5 and mAb-6 can identify A section and B section, antigen simultaneously
Epitope is located at A, B and is overlapped area, is 43-52aa；Monoclonal antibody mAb-7,8,9 only identify B section, epitope 43-70aa；mAb-
10, it 11,12,13 can identify that B section and C section, epitope are located at B, C and are overlapped area simultaneously, be 56-70aa；Monoclonal antibody mAb-
14 only identify C section, epitope 56-82aa.Testing result also display of commodity mammaglobin monoclonal antibody
MAb304 (MA512638 of PIERCE company) only identifies D section, epitope 69-93aa.
4, the Mam-A targeting of different epitope monoclonal antibodies is identified
Each one plant of representative antibodies, i.e. mAb-1, mAb-6, mAb-7, mAb-10 are chosen from above-mentioned five classes monoclonal antibody
And mAb-14, immunofluorescence experiment, distribution situation of the Mam-A that detection antibody is identified in cell, tool are carried out as primary antibody
Gymnastics is made as follows：
The breast cancer cell line ZR75-1 that reporter gene containing GFP of recovering marks, takes 24 orifice plates, prepares cell climbing sheet.Every hole
After cell density reaches 70-80%, culture medium is removed, PBS buffer solution is washed 2 times, and every hole adds 4% paraformaldehyde of 400ul, room temperature
15 minutes are fixed, PBS is washed 3 times, every time 5 minutes.Confining liquid room temperature closes 1h.5 kinds of antibody are diluted with antibody diluent.Every hole adds
Antibody after 250ul dilution, is incubated at room temperature 2h.PBS is washed 3 times, every time 5 minutes.The sheep anti mouse secondary antibody of AF594 label is diluted with antibody
Liquid is with 1：100 dilution proportions, every hole 250ul are incubated at room temperature 1h, and PBS is washed 3 times, every time 5 minutes.DAPI is with PBS with 1：1000 is dilute
It releases, every hole adds 250ul, is incubated at room temperature 15 minutes.PBS is washed 3 times, every time 5 minutes.Take out coverslip, mounting glycerol mounting, laser
The burnt lower observation of copolymerization.
ZR75-1 cell expressing green fluorescent protein (GFP) through slow-virus transfection and in cytoplasm, to observe dyeing
As a result.It is compareed using DiI dyeing (red) as cell membrane positive staining, determines film target antibody.The sheep anti mouse of AF594 label
Secondary antibody is displayed in red fluorescence, and DAPI, which is dyed, determines cell nuclear location.Observed using laser co-focusing, as a result as shown in Figure 1, with
DiI dyeing compares, and only mAb-6 is the coloring of typical cell membrane；MAb304 is Mam-A in the target cell membrane of commercialization
Monoclonal antibody, but its cell membrane is dyed and is not true to type, staining power is also weaker than mAb-6.The corresponding epitope of mAb-6 is 43-
52aa, i.e. amino acid sequence are KELLQEFIDD.
5, it interacts between BIAcore dynamics affinity analysis antigen-antibody
Using BIAcore Binding experiment detection antibody antigen interaction situation, dynamics data is shown in Table 2.Ka indicates knot
Rate constant is closed, effect combines speed between reacting macromolecular；Kd indicates dissociation rate constant, has reacted molecular complex dissociation speed
Degree；KD indicates Dissociation equilibrium constant (KD=kd/ka).Ka numerical value shows that the association rate of mAb-6 and Mam-A full-length proteins is normal
Number is significantly higher than mAb304, shows that mAb-6 and Mam-A antigen binding reaction speed is most fast in same time span.Meanwhile mAb-
6 is minimum with the Dissociation equilibrium constant of Mam-A full-length proteins, shows bond strength highest between mAb-6 and Mam-A antigen molecule.By
The above results can obtain, and mAb-6 is significantly higher than commercial antibody mAb304 to the affinity of Mam-A antigen fragment.
It interacts between 2 BIAcore dynamics affinity analysis antigen-antibody of table
By embodiment 1, we obtain the monoclonal antibody for being prepared for 14 plants of anti-human Mam-A, and screen specificity
Monoclonal antibody mAb-6, mAb-6 in conjunction with film positioning Mam-A are the coloring of typical cell membrane, are significantly better than commercialization monoclonal antibody
MAb304, in addition it is also significantly greater than commercial antibody mAb304 to the affinity of Mam-A antigen.
2 breast cancer tissue's immunohistochemistry pathological examination of embodiment
Patient with breast cancer's pathology paraffin section drying after successively immerse dimethylbenzene (I), dimethylbenzene (II), absolute alcohol,
95% alcohol, 70% alcohol.It immerses clear water after dewaxing to wash once, 5min/ times.Slice immerses in distilled water soaking and washing 5 minutes,
The box for having citric acid working solution is put into micro-wave oven high fire 3 minutes by period.Slice immerse citric acid working solution in, in it is low
Fire 13 minutes.After taking out in micro-wave oven, box cover is removed, is placed in ventilation cooling.Slice is washed 1 time, 5min/ times with TBS.It gets rid of
3%H is added dropwise after TBS2O2Room temperature is handled 15 minutes in solution wet box.Slice is washed 3 times, 3min/ times with TBS.It is dripped after getting rid of TBS liquid
Add confining liquid, 37 DEG C are closed 30 minutes.Primary antibody covers entire sample tissue after getting rid of the pre- closing that equivalent is added dropwise after deblocking liquid.Glass
Piece is put into 4 DEG C of overnight incubations in wet box.TBS is washed 3 times, 3min/ times.Slide gets rid of net dropwise addition secondary antibody working solution after drying, and is placed in wet
It is incubated for 20 minutes for 37 DEG C in box.TBS is washed 3 times, 3min/ times.It gets rid of and the anti-mouse IgG polymer working solution of HRP label is added dropwise after drying only,
It is placed in wet box and is incubated for 20 minutes for 37 DEG C.TBS is washed 3 times, 3min/ times.DAB developing solution is added dropwise after getting rid of TBS in slide, is placed in wet box
Middle room temperature is protected from light processing 10 minutes.With clear water soaking and washing 5 minutes after clear water termination reaction.Haematoxylin is immersed after slide drying
In, it dyes 3 minutes, clear water rinses 20s.Slide, which immerses in 1%HCL, handles 3s, washes 1 time.1% ammonium hydroxide is multiple about 3 minutes blue, water
It washes 1 time.80%, 95%, 100%, 100% each 2min of gradient alcohol dehydration, dimethylbenzene handle 7min.Resinene mounting.
As a result as shown in Fig. 2, cell membrane Mam-A targeting monoclonal antibody mAb-6 can specific recognition breast cancer tissue, can be with
Pathological examination for breast cancer tissue.
3 breast cancer immunofluorescent detection method of embodiment
The breast cancer cell line ZR75-1 of recovery GFP reporter gene label, takes 24 orifice plates, prepares cell climbing sheet.Every hole is thin
After born of the same parents' density reaches 70-80%, culture medium is removed, PBS buffer solution is washed 2 times, and every hole adds 400ul4% paraformaldehyde, and room temperature is fixed
15 minutes, PBS was washed 3 times, every time 5 minutes.Confining liquid room temperature closes 1h.Antibody mAb-6 antibody diluent is with 1：100 dilutions.
Antibody after every hole adds 250ul to dilute is incubated at room temperature 2h.PBS is washed 3 times, every time 5 minutes.The sheep anti mouse secondary antibody of AF594 label is used anti-
Body dilution is with 1：100 dilution proportions, every hole 250ul are incubated at room temperature 1h, and PBS is washed 3 times, every time 5 minutes.DAPI is with PBS with 1：
1000 dilutions, every hole add 250ul, are incubated at room temperature 15 minutes.PBS is washed 3 times, every time 5 minutes.Take out coverslip, mounting glycerol envelope
Piece is observed under laser co-focusing.
As a result as shown in Figure 1, cell membrane Mam-A targeting monoclonal antibody mAb-6 can specific recognition breast cancer cell, can be with
Immunofluorescence test for breast cancer cell.
4 breast cancer double-antibody sandwich elisa sero-immunity detection method of embodiment
It is coated with elisa plate with mAb-6 antibody, 100 holes μ l/, 4 DEG C overnight.3%BSA-PBS closing, 120 holes μ l/, room temperature 6
Hour, confining liquid is got rid of, drying at room temperature is spare.50 μ l test serums are added in every hole, add 50 μ l biotin labelled antibodies, fill
Divide after mixing and is incubated for 1 hour, board-washing 5 times for 37 DEG C.The 100 μ l of horseradish peroxidase for adding Avidin to mark, 37 DEG C of incubations
30min, board-washing 5 times.A, B liquid respectively adds 50 μ l, 37 DEG C of incubation 10min, 50 μ l of terminate liquid terminations, surveys OD450nm value.
It carries out double-antibody sandwich testing result to blood serum of patients with human breast carcinoma to show, 52 are sun in 60 breast cancer serums
Property, sensitivity 86.7%, and only have 2 in 60 normal control serum for the positive, specificity is 96.7%.
The capture of 5 breast cancer circulating tumor cell of embodiment and identification method
Breast cancer cell line ZR75-1 cell suspension or patient with breast cancer's anticoagulation of collection, 1000 × g are centrifuged 10min.
Carefully take at the middle and upper levels liquid be placed in EP pipe, be added and the PBS of its equivalent mixed well.Cell membrane Mam-A targeting monoclonal antibody is added
The 30 μ L of immunolipid magnetic ball of mAb-6 label, is incubated at room temperature 30min, and every 10min is mixed 1 time.EP pipe is inserted on Magneto separate frame
5min is adsorbed, inhales and abandons supernatant, PBS is washed 3 times.Add 10 μ of DAPI dyeing liquor 30 μ L and Mam-A-FITC (mAb-13) dyeing liquor
L, mixing are protected from light dyeing 15min, and PBS is washed 3 times.The resuspension of 30 μ L deionized waters is added into EP pipe, is uniformly applied to anticreep and carries glass
Piece, the fluorescence microscopy microscopic observation after drop is dry.
As a result as shown in figure 3, the immunolipid magnetic ball of cell membrane Mam-A targeting monoclonal antibody mAb-6 label can capture cream
Adenocarcinoma tumor cells.MAb-6 is labelled antibody, and mAb-13 is identification antibody.
6 mAb-6 of embodiment modifies the targeted therapy effect that PLGA carries medicine particle to breast cancer
1, mAb-6 modifies PLGA (poly lactide-glycolide acid) and carries medicine particle to the targeting of breast cancer cell
200 μ l (100mg/mL) PLGA nano particles are taken to be centrifuged, with the resuspension of EDC/NHS activation buffer, room temperature concussion is incubated
Educate 1h.With the resuspension of 200 μ l PBS buffer solution after centrifugation, 10 μ l (1mg/mL) monoclonal antibody mAb-6,4 DEG C of overnight incubations are added.PBS is slow
Fliud flushing is washed 2 times, and 200 μ l confining liquids are resuspended, and is saved backup for 4 DEG C after room temperature closing 1h, is obtained the nano particle of coupling monoclonal antibody.Claim
It takes chemotherapeutics doxorubicin hydrochloride (Dox) 0.01g to be added in PLGA oil phase to mix, prepares the load medicine particle of coupling monoclonal antibody
(mAb-NP-Dox)。
ZR75-1 cell is passed to 24 orifice plates and normally cultivates, to cell density up to 60% or so, be separately added into independent Dox,
PLGA carries medicine particle (NP-Dox) and mAb-6 modification PLGA carries medicine particle (mAb-NP-Dox), 37 DEG C of incubation 1h, nuclei dyeing
Color, fluorescence microscopy is under the microscope.Load medicine particle is red fluorescence, and nucleus is blue-fluorescence.
As a result as shown in figure 4, breast cancer cell cannot be targeted by being individually added into Dox, it is visible that PLGA carries medicine particle (NP-Dox)
It is adhered to cell to a small amount of Dox, and mAb-6 modification PLGA carries medicine particle (mAb-NP-Dox) visible a large amount of Dox and is adhered to cell,
Illustrate that mAb-6 antibody modification PLGA carries medicine particle and has good targeting, it can be by therapeutic agent targeted delivery to breast cancer
Cell.
2, mAb-6 modifies PLGA and carries medicine particle to the targeted inhibition effect of Cells Proliferation of Human Breast Cancer
The PLGA for carrying chemotherapeutics Dox is carried into medicine particle (NP-Dox) respectively and mAb-6 modification PLGA carries medicine particle
(mAb-NP-Dox) normal incubation medium is replaced after being incubated for 12h altogether with ZR75-1 breast cancer cell, is detected after 48h.Ki67 exempts from
Epidemic disease groupization detects cell Proliferation, and positive cell rate statistics is shown in Fig. 5.It can be obtained from the figure that mAb-6 modification PLGA carries medicine particle group (mAb-
NP-Dox) PLGA that positive cell rate does not carry out monoclonal antibody modification substantially less than carries medicine particle (NP-Dox) group, illustrates that mAb-6 is modified
It is more significant to the targeted inhibition effect of Cells Proliferation of Human Breast Cancer that PLGA carries medicine particle.
The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；Although referring to aforementioned each reality
Applying example, invention is explained in detail, those skilled in the art should understand that：It still can be to aforementioned each
Technical solution documented by embodiment is modified, or equivalent substitution of some or all of the technical features；And
These are modified or replaceed, the range for technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution.
Sequence table
<110>Military medical research institute of PLA Academy of Military Sciences
<120>A kind of mammaglobin Staphylococal Protein A epitope and its application
<130> 1
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
Claims (10)
1. a kind of mammaglobin Staphylococal Protein A epitope, amino acid sequence is as shown in sequence table SEQ ID NO.1.
2. application of the mammaglobin Staphylococal Protein A epitope described in claim 1 in the antibody for preparing mammaglobin A.
3. application according to claim 2, the antibody is monoclonal antibody.
4. a kind of monoclonal antibody of mammaglobin A, which is characterized in that it identifies epitope described in claim 1.
5. epitope described in claim 1 or monoclonal antibody as claimed in claim 3 are in preparation prevention or treatment mammary gland
The reagent of globin A abnormal expression associated diseases or the purposes in drug.
6. epitope described in claim 1 or monoclonal antibody as claimed in claim 3 are in preparation detection, prevention or treatment
Purposes in breast cancer reagent or drug.
7. purposes according to claim 6, the reagent be immunohistochemistry detection reagent, immunofluorescence detection agent,
ELISA detection reagent, breast cancer circulating tumor cell capture reagent.
8. a kind of target composition for treating breast cancer, which is characterized in that include monoclonal antibody as claimed in claim 4.
9. target composition according to claim 8, which is characterized in that further include azithromycin dihydrochloride.
10. target composition according to claim 9, which is characterized in that further include PLGA nano particle, the PLGA receives
Rice grain is in conjunction with monoclonal antibody as claimed in claim 4.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113960302A (en) * | 2021-09-29 | 2022-01-21 | 中国人民解放军军事科学院军事医学研究院 | Kidney toxicity detection method based on high content technology and application thereof |
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| CN113960302A (en) * | 2021-09-29 | 2022-01-21 | 中国人民解放军军事科学院军事医学研究院 | Kidney toxicity detection method based on high content technology and application thereof |
| CN113960302B (en) * | 2021-09-29 | 2024-04-23 | 中国人民解放军军事科学院军事医学研究院 | Nephrotoxicity detection method based on high content technology and application thereof |
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