CN105218643A - A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation - Google Patents
A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation Download PDFInfo
- Publication number
- CN105218643A CN105218643A CN201510718349.3A CN201510718349A CN105218643A CN 105218643 A CN105218643 A CN 105218643A CN 201510718349 A CN201510718349 A CN 201510718349A CN 105218643 A CN105218643 A CN 105218643A
- Authority
- CN
- China
- Prior art keywords
- antibacterial peptide
- lactobacterium casei
- antibody
- synthesis
- polypeptide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention discloses a kind of lactobacterium casei antibacterial peptide and many anti-preparation methods thereof.Antibacterial peptide of the present invention is template engineer and a kind of micromolecule polypeptide of synthesis with natural fine rhzomorph (antibacterial peptide) product of lactobacterium casei fermentation, comprise 16 amino-acid residues, its aminoacid sequence is: MDSLKTLLVANRGEIV, and its polyclonal antibody preparation step is as follows: the Design and synthesis of (1) lactobacterium casei antibacterial peptide; (2) improvement on synthesis and carrier proteins are cross-linked; (3) animal immune is carried out with lactobacterium casei polypeptide immunogen; (5) in the animal body after immunity, get blood, collect, be separated the serum obtained containing antibody, antibody purification, namely obtains the antibody of anti-lactobacterium casei polypeptide.Lactobacterium casei antibacterial peptide of the present invention has that molecular weight is little, the simple feature of synthetic.Have the feature of the good and high-titer of specificity with the inventive method Dispersal risk, the application producing the detection field of bacteriocin (antibacterial peptide) for lactobacterium casei fermentation provides important tool.
Description
Technical field
The invention belongs to biological technical field, relate to the preparation method of a peptide species and antibody thereof, especially for for lactobacterium casei antibacterial peptide and the polyclonal antibody of anti-lactobacterium casei antibacterial peptide and preparation method thereof.
Background technology
Along with people's life and the improving constantly of health level, the demand of people to green food is increasing, and Consciousness of food security is also more and more higher.And apply the feeding antibiotic of over half a century in the whole world, although at raising efficiency of feed utilization, improve the output of live-stock product, increase livestock breed aquatics benefit etc. in played great function; But, along with antibiotic widespread use is even abused, not only cause the generation of endurance strain, also can cause the drug residue of livestock product, bring food safety hidden danger.Therefore, research and development safety, efficient novel feeding antibiotic substitute products become focus.
Along with going deep into of research, Chinese scholars finds that antibacterial peptide (antimicrobialpeptides, AMPs) has very excellent characteristic: as suppressed multiple pathogenic microorganisms, have good selectivity sterilization effect; Some proteolytic enzyme (as trypsinase, Alphachymdean Catarasce etc.) in easy digested road degrade, do not produce accumulative accumulation in animal body; Have no side effect, noresidue, without resistance, avoid the threat that agricultural antibiotic is produced human body by the transmission of food chain, simultaneously also free from environmental pollution; Antibacterial peptide has the beneficial effect of similar antibiotic feed additive in a word, meets the needs that livestock product safety is produced, is adapted at using in fodder production, has the potential quality as fodder additives of new generation.But up to the present, still the caning be counted on one's fingers at feed industry as fodder additives large-scale application of antibacterial peptide.
In addition, in China, lactobacterium casei industry development is still in the starting stage, domestic lactobacterium casei industry production business or retailer's sales volume lower, its species L. casei used in enormous quantities and starter are mostly from famous foreign manufacturer, and therefore lactobacterium casei industry also has larger development space; Current lactobacterium casei is at food, particularly there is certain application in milk-product, but the research added in feed is also little, and domestic scholar is to the process study of lactobacterium casei bacteriocinogeny (having another name called antibacterial peptide) and detect also in the starting stage, and commercially produced product is also very limited.
At present, relevant lactobacillus casei bacteriocin (antibacterial peptide) patent application is as follows: application number: 201010277557.1, denomination of invention " a kind of lactobacillus casei bacteriocin and the application in feed thereof ", this patent relates to a kind of lactobacillus casei bacteriocin and the application in feed thereof, and the lactobacterium casei (Lactobacilluscasei) specifically disclosed described in a strain lactobacterium casei, preservation name is called lactobacterium casei LyT-7, its depositary institution: China typical culture collection center, preserving number is: CCTCCNO:M2010197, and produce the method for bacteriocin, with the application of bacteriocin crude extract in feed, provide a kind of lactobacillus casei bacteriocin, it is stable to heat, acid, can be easily degraded by proteases, and antimicrobial spectrum is wide, has the prospect as fodder additives application.But do not relate to for lactobacterium casei fermentation manufacturing technique with as the large-scale production process of fodder additives, also do not relate to bacterial detection element (antibacterial peptide) rapid detection.
Application number: 201310162105.2, denomination of invention is " application of a kind of lactobacterium casei fermented liquid in feed ", the application of fermented liquid in feed of a kind of lactobacterium casei LyT-7 is disclosed, the lactobacterium casei fermented liquid that the large scale fermentation specifically disclosing this bacterial strain obtains, the fodder additives that the rape cake by spraying dry and after adding detoxification is made.But the accurately detection fast of the bacteriocin (antibacterial peptide) for concrete fermented liquid, does not have further disclosure; Therefore in order to adapt to large scale fermentation produce in for bacteriocin (antibacterial peptide) tire in the urgent need to, the technique study that a kind of rapid detection bacteriocin (antibacterial peptide) is tired is necessary.
And immunologic detection method is because of high specificity, the advantage such as highly sensitive and easy and simple to handle and enjoy favor.But at present for the antibody of the institute of lactobacillus casei bacteriocin (antibacterial peptide), market there is no commercially produced product.Therefore, how to obtain high specificity, antibody that potency ratio is high just become problem demanding prompt solution in research, but by the physico-chemical property of natural fine rhzomorph itself, high purity object bacteriocin acquisition not a duck soup, thus have impact on the acquisition of corresponding polyclonal antibody; Based on above reason, we intend synthetic lactobacterium casei antibacterial peptide, and the lactobacterium casei antibacterial peptide polyclonal antibody that preparation is relevant, to so that the rapid detection of lactobacterium casei antibacterial peptide basis is provided.
Summary of the invention
One object of the present invention is the synthesis providing a kind of lactobacterium casei antibacterial peptide.The one that the present invention utilizes this bacterial strain to produce, to heat, ph stability, has the natural fine rhzomorph (having another name called antibacterial peptide) of broad-spectrum antibacterial, after structural analysis is carried out to this bacteriocin, and synthetic.
The depositary institution of this lactobacterium casei is: China typical culture collection center, preservation address is: China. Wuhan. and Wuhan University, the preservation time is: 2010 08 month No. 09, preserving number is: CCTCCNO:M2010197, and its Classification And Nomenclature is: lactobacterium casei lyT-7 (LactobacilluscaseilyT-7).
Object of the present invention is realized by following technology:
Step one: described natural fine rhzomorph obtains in accordance with the following methods: for cultivating the substratum of lactobacterium casei lyT-7 be: glucose 2%, peptone 1%, yeast extract paste 0.5%.Described culture temperature is 30 DEG C, quiescent culture 48h; The fermented liquid centrifuging and taking supernatant obtained.
Step 2: add ammonium sulfate to whole saturation ratio 50% to this supernatant liquor and saltout; Then centrifuging and taking precipitation, throw out being loaded flow is dialyse in pure water in the dialysis tubing of 1000, and concentrates with polyoxyethylene glycol, obtains bacteriocin crude extract; This bacteriocin crude extract is carried out further through SephadexG-100 gel column purifying; The bacteriocin sample collected is carried out RP-HPLC separation, obtains the final bacteriocin having bacteriostatic activity.
Step 3: adopt mass spectrograph to carry out structural analysis to the bacteriocin finally obtained.This bacteriocin comprises 16 amino-acid residues, and its total order is classified as: MDSLKTLLVANRGEIV(N-C).
Step 4: adopt solid-state chemical reaction method method, with polypeptide automatic DNA synthesizer DNA synthesis imitation cheeses Bacterium lacticum antibacterial peptide.When synthesizing described peptide sequence, hold increase halfcystine, to facilitate and carrier protein couplet at its C.
Step 5: use high performance liquid chromatography to carry out purity testing the polypeptide of synthesis, and use electrospray mass spectrometry to identify improvement on synthesis, complete the preparation of polypeptide.
Another object of the present invention is to provide that a species specificity is good, purity is high, can with polypeptide polyclonal antibody of natural lactobacillus casei bacteriocin (antibacterial peptide) specific recognition and preparation method thereof.
Described lactobacterium casei antibacterial peptide polyclonal antibody is achieved through the following technical solutions:
Step one: the antibacterial peptide after purifying and keyhole limpet hemocyanin (KeyholeLimpetHemocyanin, KLH) coupling are prepared immunogen, prepares coating antigen with bovine serum albumin (Bovinealbumin, BSA) coupling.
Step 2: the immunizing antigen obtained mixes with Freund's complete adjuvant equal-volume, after fully emulsified, carries out neck dorsal sc multi-point injection to New Zealand white rabbit, and after initial immunity, immunity in every 2 weeks 1 time, carries out 3 booster immunizations.
Step 3: last booster immunization is after 10 days, and put to death by rabbit, carotid artery gets blood.Collect, be separated the serum obtained containing rabbit anti-lactobacterium casei antibacterial peptide antibody.
Step 4: by antiserum(antisera) by carrying out peptide affinity purification with affinity chromatography, obtain lactobacterium casei antibacterial peptide antibody.
Step 5: ELISA bioactivity is carried out to lactobacterium casei antibacterial peptide antibody.
Accompanying drawing explanation
Fig. 1: the HPLC detected result showing the purity check of lactobacterium casei antibacterial peptide synthesis.Wherein " * " represents object peak, and ordinate zou represents the absorption value of elution peak at 220nm, and X-coordinate represents elution time.
Fig. 2: the mass spectrum showing the Primary Structure Analysis of lactobacterium casei antibacterial peptide synthesis.Wherein ordinate zou represents relative abundance (%), and X-coordinate represents mass-to-charge ratio (m/z).
Embodiment
The following examples further illustrate essentiality content of the present invention, should be understood that these embodiments only for explaining the present invention, and are not used in and limit the scope of the invention.
Embodiment 1: the design of lactobacterium casei antibacterial peptide and synthesis
1, lactobacterium casei (Lactobacilluscasei) lyT-7 produces the isolation and purification of natural fine rhzomorph
Described natural fine rhzomorph obtains in accordance with the following methods: accessed in MRS substratum by lactobacterium casei lyT-7, its main component is: glucose 2%, peptone 1%, yeast extract paste 0.5%.Described culture temperature is 30 DEG C, static gas wave refrigerator 48h; The fermented liquid centrifuging and taking supernatant obtained, add ammonium sulfate to whole saturation ratio 50% to this supernatant liquor and saltout, centrifuging and taking precipitates; Throw out being loaded flow is dialyse in pure water in the dialysis tubing of 1000, and concentrates with polyoxyethylene glycol, obtains bacteriocin crude extract; This bacteriocin crude extract is carried out further through SephadexG-100 gel column purifying; The bacteriocin sample collected is carried out RP-HPLC separation, obtains the final bacteriocin having bacteriostatic activity; Adopt mass spectrograph to carry out structural analysis, the aminoacid sequence analysis of MS-MS spectrum being obtained to this bacteriocin is: MDSLKTLLVANRGEIV.
2, the synthesis of lactobacterium casei antibacterial peptide
According to aminoacid sequence described above, synthesize with automatic Peptide synthesizer, method is solid-state chemical reaction method method.For ease of being cross-linked with carrier proteins, increasing the immunogenicity of polypeptide, increase a halfcystine C at the C-terminal of aforementioned polypeptides, therefore peptide sequence finally to be synthesized being MDSLKTLLVANRGEIVC.The polypeptide high performance liquid chromatograph (HPLC) of synthesis detects purity, and purity is higher, and assorted peak is few, and this synthetic antibacterial peptide purity is 93.7% as calculated, meets immune purity requirement.Synthetic antibacterial peptide MS mass spectrograph detects, mass-to-charge ratio (m/z) [M+2H] of polypeptide
2+the molecular weight of quasi-molecular ions gained is 1862.25, basically identical with theoretical molecular, proves the accuracy of synthetic antimicrobial peptide sequence from molecular weight angle.
Embodiment 2: prepare immunogenic method
By antibacterial peptide and keyhole limpet hemocyanin (Keyholelimpethemocyanin, KLH) coupling prepares immunogen, and coupling agent is 4-(N-maleimidomehyl) hexanaphthene-1-carboxylic acid sulfonic group succinimide ester sodium salt (Sulfo-SMCC).Polypeptide and bovine serum albumin (Bullserumalbumin, BSA) coupling (coupling agent is glutaraldehyde) simultaneously, as detectable antigens.How the key of anti-preparation is the synthesis of immunizing antigen, and immunizing antigen needs purity good, and can keep the chemical structure of antibacterial peptide, is conducive to the generation of follow-up animal internal antibody.
Embodiment 3: the process of animal immune
Choose of the right age New Zealand white rabbit as immune animal, ear venous blood sampling 2-3mL before immunity, for the negative control that follow-up ELISA detects.During first immunisation, being diluted by immunizing antigen with PBS is 1mg/mL, gets 1mL antigen, mixes with the freund's adjuvant of equivalent, fully mixes emulsification (inspection emulsification degree: by an emulsification antigen liquid instillation physiological saline, if do not scatter, show to reach requirement).Rabbit neck dorsal sc multiple spot (at least 8 point) is injected.After two weeks, carry out first time booster immunization, full freund's adjuvant emulsification of cannoing be used up, injection site is identical with initial immunity with dosage.After this booster immunization of same operation was carried out every 2 weeks, totally 3 times, front and back.Last booster immunization, after 10 days, adopts the mode of carotid artery bloodletting to collect rabbit blood, prepares serum.
Embodiment 4: affinity column antibody purification
1mg polypeptide is connected on the SulfolinkResin of 1mL activation, prepares antigen affinity column; Affinity column 10 times of column volume PBS balance, and flow to end solution; Rabbit anteserum, through 0.45um aperture membrane filtration, adds rabbit anteserum to antigen affinity column, and antigen affinity column crossed by serum, flows to end solution; After PBS buffer solution elution 10 column volumes, with 5mL0.1M glycine (pH2.7) wash-out, be in charge of collection elutriant, often pipe 1mL; The elutriant collected detects the absorbancy at 280nm place, the component merging that absorbancy is greater than 1.0, to PBS dialysis, obtains resisting of purifying more.
Embodiment 5: indirect elisa method measures tiring of antibody
Being diluted by detectable antigens CBS is 1ug/mL, adds in enzyme plate, every hole 100uL, 4 DEG C of bag quilts that spend the night; Next day, discarded by coating buffer, every hole adds 200ul confining liquid (5% skim-milk), 37 DEG C of closed 1.5h; Discard confining liquid, do different concns dilution with lactobacterium casei antibacterial peptide antibody, 1:1250,1:2500,1:5000,1:10000,1:20000,1:40000,1:80000, and negative serum adds enzyme mark hole, every hole 100uL, hatches 1h for 37 DEG C.Add the goat-anti rabbit of horseradish peroxidase mark two resist, and every hole 100uL, adds enzyme plate, hatches 30min for 37 DEG C.The TMB adding every hole 100uL carries out color reaction, and 15min is placed in 37 DEG C of dark places.Measure the absorbancy of 450nm wavelength after vitriol oil termination reaction, calculating antibody is tired.Experimental result is as shown in the table.When the ratio with negative serum is greater than 2.1, calculating antibody is tired.After testing, tiring of the rabbit polyclonal antibody after purifying reaches 1:80000.
SEQUENCELISTING
<110> Sichuan University
The synthesis of a <120> lactobacterium casei antibacterial peptide and antibody preparation
<130>2015
<160>1
<170>PatentInversion3.3
<210>1
<211>16
<212>PRT
<213> artificial sequence
<400>1
MetAspSerLeuLysThrLeuLeuValAlaAsnArgGlyGluIleVal
151015
Claims (10)
1. the synthesis of a lactobacterium casei antibacterial peptide and antibody preparation, described lactobacterium casei (Lactobacilluscasei) preservation name is called lactobacterium casei LyT-7, it is characterized in that: the depositary institution in lactobacterium casei fermented liquid: China typical culture collection center, preserving number is: CCTCCNO:M2010197, it is characterized in that: be a kind of antibacterial peptide of template synthetic with natural fine rhzomorph (antibacterial peptide) product in lactobacterium casei LyT-7 fermented liquid, and prepare its antibody for this antibacterial peptide.
2. antibacterial peptide according to claim 1, its aminoacid sequence is MDSLKTLLVANRGEIV.
3. lactobacterium casei antibacterial peptide sequence according to claim 2, adopts solid-state chemical reaction method method, synthetic modified polypeptide.
4. the preparation method of antibacterial peptide according to claim 3, is characterized in that: described modified polypeptide is that the sequence carboxyl terminal of claim 2 additionally increases a cysteine residues, is beneficial to and carrier protein couplet.
5. carrier proteins described in is keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA), and described polypeptide obtains immunogen with described carrier protein KLH respectively, with BSA coupling obtained detect former.
6. the how anti-preparation method of a rabbit anti-lactobacterium casei antibacterial peptide, it is characterized in that: by the immunogen polypeptide of synthesis and carrier proteins being cross-linked to form conjugated protein, use immunogen immune animal, in the animal body after immunity, get blood prepare antiserum(antisera), separation and purification antibody from serum.
7. preparation method for antibody according to claim 5, is characterized in that: described animal is New Zealand white rabbit (at 3 months monthly age, body weight is about 2kg).
8. after the lactobacterium casei antibacterial peptide immunogen described in and immunological adjuvant mixing and emulsifying, carry out neck dorsal sc multi-point injection to animal, immunity in every 2 weeks 1 time after initial immunity, booster immunization number of times is 3 times.
9. last booster immunization is after 10 days, gets blood and obtains antiserum(antisera).
10. preparation method for antibody according to claim 5, is characterized in that: right wants the antiserum(antisera) obtained in 6 can obtain highly purified antibody from antiserum(antisera) after affinity purification.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510718349.3A CN105218643A (en) | 2015-10-30 | 2015-10-30 | A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510718349.3A CN105218643A (en) | 2015-10-30 | 2015-10-30 | A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105218643A true CN105218643A (en) | 2016-01-06 |
Family
ID=54987971
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510718349.3A Pending CN105218643A (en) | 2015-10-30 | 2015-10-30 | A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105218643A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105622745A (en) * | 2016-01-22 | 2016-06-01 | 深圳市新产业生物医学工程股份有限公司 | Derivative of amino acid fragment of angiotensin II, angiotensin II antigen and preparation method and application of angiotensin II antigen |
| CN105646668A (en) * | 2016-01-22 | 2016-06-08 | 深圳市新产业生物医学工程股份有限公司 | Derivative of angiotensin I amino acid segment, angiotensin I antigen and preparation and application of angiotensin I antigen |
| CN106018823A (en) * | 2016-05-19 | 2016-10-12 | 四川大学 | Method for rapidly detecting antibacterial peptides in lactobacillus casei fermentation process and final products |
| CN109030821A (en) * | 2018-07-19 | 2018-12-18 | 北京农学院 | A kind of antibody, kit and its application method of lactobacillus plantarum bacteriocin |
| CN110105433A (en) * | 2018-12-18 | 2019-08-09 | 天津科技大学 | The application of novel lactic acid bacteria antibacterial peptide and high efficient expression and antibacterial anticancer activity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101451158A (en) * | 2007-12-04 | 2009-06-10 | 黑龙江大学 | Separation and purification method of antibiotic peptides produced by Lactobacillus paracasei |
| CN102061271A (en) * | 2010-09-09 | 2011-05-18 | 四川大学 | Lactobacillus casei bacteriocin and use thereof in feed |
-
2015
- 2015-10-30 CN CN201510718349.3A patent/CN105218643A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101451158A (en) * | 2007-12-04 | 2009-06-10 | 黑龙江大学 | Separation and purification method of antibiotic peptides produced by Lactobacillus paracasei |
| CN102061271A (en) * | 2010-09-09 | 2011-05-18 | 四川大学 | Lactobacillus casei bacteriocin and use thereof in feed |
Non-Patent Citations (4)
| Title |
|---|
| 张贵君等: "《常用中药生物鉴定》", 31 January 2006 * |
| 王晶等: "Sao蛋白多克隆抗体制备鉴定及促全血杀菌作用", 《中国公共卫生》 * |
| 贾士儒: "《生物防腐剂》", 30 September 2009 * |
| 贾连群等: "《现代基础医学理论与技术进展》", 31 August 2015 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105622745A (en) * | 2016-01-22 | 2016-06-01 | 深圳市新产业生物医学工程股份有限公司 | Derivative of amino acid fragment of angiotensin II, angiotensin II antigen and preparation method and application of angiotensin II antigen |
| CN105646668A (en) * | 2016-01-22 | 2016-06-08 | 深圳市新产业生物医学工程股份有限公司 | Derivative of angiotensin I amino acid segment, angiotensin I antigen and preparation and application of angiotensin I antigen |
| CN106018823A (en) * | 2016-05-19 | 2016-10-12 | 四川大学 | Method for rapidly detecting antibacterial peptides in lactobacillus casei fermentation process and final products |
| CN109030821A (en) * | 2018-07-19 | 2018-12-18 | 北京农学院 | A kind of antibody, kit and its application method of lactobacillus plantarum bacteriocin |
| CN109030821B (en) * | 2018-07-19 | 2021-07-20 | 北京农学院 | Antibody and kit of lactobacillus plantarum bacteriocin and use method of antibody and kit |
| CN110105433A (en) * | 2018-12-18 | 2019-08-09 | 天津科技大学 | The application of novel lactic acid bacteria antibacterial peptide and high efficient expression and antibacterial anticancer activity |
| CN110105433B (en) * | 2018-12-18 | 2022-06-14 | 天津科技大学 | Lactic acid bacteria antibacterial peptide and application of high-efficiency expression and antibacterial and anticancer activity |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105218643A (en) | A kind of synthesis of lactobacterium casei antibacterial peptide and antibody preparation | |
| CN102747042B (en) | Hybridoma cell line 10G4 and anti-alfatoxin total (B1, B2, G1 and G2) monoclonal antibody produced by the same | |
| CN104497137B (en) | The general monoclonal antibody of African swine fever virus strain and preparation method and application | |
| CN107656066B (en) | A kind of duck tembusu virus E protein truncated protein and application | |
| CN103033626B (en) | Colloidal gold strip for TGEV antibody and PEDV antibody | |
| CN108414762B (en) | Grass carp IgM monoclonal antibody, preparation method and application thereof | |
| CN103163299A (en) | Avian leukosis double-antibody sandwich enzyme-linked immuno sorbent assay (ELISA) antigen detection kit | |
| CN103436496A (en) | Anti-mycoplasma bovis monoclonal antibody, hybridoma cell strain secreting monoclonal antibody and application | |
| CN101329339B (en) | Method for testing cattle immune globulin G and special-purpose enzyme-linked immunologic reagent kit thereof | |
| CN103992988A (en) | Hybridoma cell line and anti-canine distemper virus N protein monoclonal antibody produced through hybridoma cell line | |
| San-Blas et al. | Paracoccidioidomycosis | |
| CN102690787B (en) | Hybridoma cells capable of secreting an anti human [beta]-actin protein monoclonal antibody and preparation method thereof | |
| CN103033631B (en) | Test kit for qualitatively detecting fish vitellogenin by utilizing lipovitellin antibody | |
| CN105087498A (en) | TCMTB monoclonal antibody hybridoma cell strain and application thereof | |
| CN103665139A (en) | Adipocyte fatty-acid binding protein and preparation and application of monoclonal antibody of adipocyte fatty-acid binding protein | |
| CN103725697A (en) | Chemically synthesized staphylococcus aureus surface protein FnBPA gene fragment and expression and application thereof | |
| CN103204936A (en) | Preparation method for polyclonal antibody to salmonella effect protein SopB | |
| Conway de Macario et al. | Six antigenic determinants in the surface layer of the archaebacterium Methanococcus vannielii revealed by monoclonal antibodies. | |
| CN110261606A (en) | A kind of C. perfringens beta toxin Anti-HBV permanence detection method | |
| CN103288954A (en) | Monoclonal antibody with rock bream iridovirus ORF049L recombinant proteins and preparation method thereof | |
| CN102532310B (en) | Monoclonal antibody against grass carp reovirus VP5 protein and application of monoclonal antibody | |
| CN103342740B (en) | A kind of blocking ELISA method for detecting fowl HEV specific antibody | |
| CN103450358A (en) | Swine GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) protein antibody and preparation method and application thereof | |
| CN110590561A (en) | Herbicidal ether hapten, artificial antigen and antibody as well as preparation method and application thereof | |
| CN110713985B (en) | Hybridoma cell strain 5H4 secreting monoclonal antibody against CLas membrane protein Cmp1, monoclonal antibody and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160106 |