CN102432685A - Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres - Google Patents
Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres Download PDFInfo
- Publication number
- CN102432685A CN102432685A CN2011103395428A CN201110339542A CN102432685A CN 102432685 A CN102432685 A CN 102432685A CN 2011103395428 A CN2011103395428 A CN 2011103395428A CN 201110339542 A CN201110339542 A CN 201110339542A CN 102432685 A CN102432685 A CN 102432685A
- Authority
- CN
- China
- Prior art keywords
- mbfgf
- mouse
- immune latex
- immune
- gene
- 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
Images
Abstract
The invention relates to immune latex microspheres for detecting a recombined mouse basic fibroblast growth factor (bFGF) and a preparation method and application of the immune latex microspheres. The immune latex microspheres are coated by an anti-mbFGF protein polyclonal antibody, and the particle size of the microspheres is 50nm. The preparation method comprises the following steps of: re-modifying and reconstructing a full-length mbFGF gene expression sequence according to a priority codon expressed by procaryotic cells; inserting the gene sequence into a glutathione-S-transferase gene fusion expression vector through restriction enzyme digestion, and performing bacterial expression; obtaining an mbFGF protein through affinity chromatographic column purification and thrombin online digestion; fusing an adjuvant with an antigen, and then immunizing a rabbit; separating and purifying to obtain the anti-mbFGF polyclonal antibody; and coupling the antibody to activated microspheres through a chemical coupling method to obtain the immune latex microspheres for detecting the mbFGF protein. The application of the immune latex microspheres is as follows: through an enzyme linked immunosorbent assay method, an mbFGF protein sample is detected, the lowest limit of detection is 60pg/ml of the mbFGF protein, and a related coefficient R2 is equal to 0.96. The immune latex microspheres disclosed by the invention have the advantages of high detection sensibility, simpleness in operation and strong repeatability.
Description
Technical field
The present invention relates to a kind of immune latex microsphere that detects the recombined small-mouse Prostatropin.
Background technology
(basic fibroblast growth factor is by hypophysis and hypothalamus excretory polypeptide bFGF) to Prostatropin, extensively is present in nervous tissue, inoblast and the scavenger cell.As a kind of wide spectrum mitogen, it has extremely strong biological activity: 1. to deriving from mesoderm and neuroectodermal various cell short proliferation function, chemotaxis and short cell migration campaign effect are arranged all; 2. can promote the section angiogenesis, improve the nutritional status of microcirculation and tissue, promote inoblast and myoblastic propagation simultaneously, make the granulation ramp; 3. strengthen coming from mesoblastic activity of immune cells, improve the anti-infection ability of section.Research shows that bFGF participates in the processes of wound repair of multiple tissue in body, is one of wound healing factor important in the body.The high low energy of the content of bFGF directly reflects the vasculogenesis level in the wound repair after the wound, for detecting and monitor trauma repair and organization healing situation etc. foundation is provided.
Clinically, Prostatropin can be used for treating cranium brain, spinal cord, peripheral nerve injury, poisons, and infects; Hematencephalon, cerebral infarction, encephalatrophy, cerebral dysplasia, keratohelcosis, neural heariing loss, facial paralysis; Mucous membrane, skin injury, burn, chronic ulcer, soft tissue injury; Visceral lesion, fracture, some endocrinopathys, the replantation of a severed limb and various postoperative wound healings etc.BFGF can promote wound healing, shortens the course of disease greatly, improves prognosis, but it is few to send out the bFGF content detecting method for damage surface of a wound Central Plains so far, and detection sensitivity, detectability and immunity from interference etc. are in urgent need to be improved.At present, about the RR of bFGF immunoassay technology and related patent U.S. Patent No. technology both at home and abroad report seldom, press for exploitation efficiently, novel immunoassay technology and product accurately and rapidly.
Summary of the invention
The objective of the invention is to above-mentioned present situation; Aim to provide that a kind of to be used for detection sensitivity higher; And simple to operate, repeated strong, lowest detection is limited to the immune latex microsphere of the detection recombined small-mouse Prostatropin of mbFGF protein 60 pg/ml.
The implementation of the object of the invention does; One detects the immune latex microsphere of recombined small-mouse Prostatropin; Be to use the monodisperse carboxyl polystyrene microsphere to be substrate material; With the immune latex microsphere that the anti-mbFGF polyclonal antibody of carboxyl-amino chemical coupling method coupling obtains, be the microballoon that encapsulates by anti-mbFGF protein polyclone antibody, microsphere particle size 50nm.
Detect the preparation method of the immune latex microsphere of recombined small-mouse Prostatropin, concrete steps are following:
(1) modifies and makes up total length mouse bFGF expressed sequence again according to the preferential codon of procaryotic cell expression;
(2) mouse bFGF gene fragment is gone in the glutathione-S-transferase gst gene fusion expression vector through the restriction enzyme digestion cleft cutting, make up the bacterium fusion expression vector;
Said glutathione-s-transferase gene fusion expression vector is pGEX-4T-3, and the fusion expression vector that makes up after accomplishing is pGEX-4T-3-mbFGF;
(3) the bacterium fusion expression vector is carried out bacterial expression after, utilize the affinity chromatography column purification to obtain the GST-mbFGF fusion rotein; Said affinity chromatographic column is Glutathione Sepharose 4B;
(4) utilize zymoplasm at post cutting GST-mbFGF fusion rotein, eluate obtains mouse bFGF after affinity chromatographic column is removed zymoplasm; Affinity chromatographic column is Benzamidine Sepharose 4B Fast Flow:
(5) with immunize rabbit behind the fused antigen of freund's adjuvant, obtain the anti-mbFGF polyclonal antibody of rabbit through separation and purification; Said antigen is mbFGF albumen;
(6),, obtain detecting the immune latex microsphere of recombined small-mouse Prostatropin through carboxyl-amino chemical coupling method coupling activation microballoon with the anti-mbFGF polyclonal antibody of the resulting rabbit of step (5);
Said microballoon is the monodisperse carboxyl polystyrene microsphere, big or small 50nm, and it is carboxyl that group is carried on the surface,
The activation of said microballoon is at first to use acvator carbodiimide activation microballoon, adds protective material Sulfo-NHS then and makes microballoon form stable active ester midbody, in order to connecting antibody molecule.
Detect the recombined small-mouse Prostatropin immune latex microsphere application, through ELIASA, adopt enzyme-linked immunosorbent assay, detect the bFGF of mbFGF sample.
It is substrate material that the present invention adopts the monodisperse carboxyl polystyrene microsphere, obtains immune latex microsphere with carboxyl-anti-mbFGF polyclonal antibody of amino chemical coupling method coupling, and it is simple, easy to operate to draw materials; The preparation method is careful rigorous, and repeatability is high; Prepared immune latex microsphere particle is little, and particle diameter is even, is recombined small-mouse Prostatropin 60pg/ml through enzyme-linked immunosorbent assay (ELISA) LDL, coefficient R
2=0.96.
Description of drawings
Fig. 1 is the carboxylic polystyrene microsphere stereoscan photograph,
Fig. 2 is the former sequence of mbFGF gene,
Fig. 3 is that bacterium colony PCR checking mbFGF gene inserts the fragment electrophoresis,
Fig. 4 is a plasmid Xba I restriction enzyme digestion and electrophoresis,
Fig. 5 is the order-checking of mbFGF expression vector,
Fig. 6 is that mbFGF protein SDS-PAGE (A) and Western Blotting (B) detect,
Fig. 7 is antiserum titre mensuration figure,
Fig. 8 is that the ELISA method detects the mbFGF pictorial diagram,
Fig. 9 is that the ELISA method detects mbFGF albumen result.
Embodiment
A kind of immune latex microsphere that detects the recombined small-mouse Prostatropin of the present invention is to use the monodisperse carboxyl polystyrene microsphere to be substrate material, obtains immune latex microsphere as shown in Figure 1 with the anti-mbFGF polyclonal antibody of carboxyl-amino chemical coupling method coupling.The immunity latex microsphere is encapsulated by anti-mbFGF protein polyclone antibody, microsphere particle size 50nm.
A kind of preparation method who detects the immune latex microsphere of recombined small-mouse Prostatropin is detailed below, and concrete steps are following:
1, modify and make up total length mouse bFGF expressed sequence again according to the preferential codon of procaryotic cell expression, its concrete steps are:
(1) design total length mbFGF gene order is modified according to the preferential codon of procaryotic cell expression again, adopts full gene synthesis technology to make up total length mbFGF gene, and totally 462 pairs of bases are cloned into carrier pGEM-T Easy, and the result is as shown in Figure 2.
(2) exactness of inserting through bacterium colony PCR checking mbFGF gene segment, the result is as shown in Figure 3.
(3) will clone mbFGF gene bacterium and implement amplification cultivation, and extract and carry out that enzyme is cut and agarose electrophoretic analysis behind the plasmid, the result is as shown in Figure 4.
(5) dna sequencing guarantees that the full sequence 100% of gene is accurate, and the result is as shown in Figure 5.
2, mouse bFGF gene fragment is gone among the glutathione-S-transferase gst gene fusion expression vector pGEX-4T-3 through the restriction enzyme digestion cleft cutting, make up bacterium fusion expression vector pGEX-4T-3-mbFGF, concrete operation steps is following:
(1) the mbFGF gene fragment is gone in the pGEX-4T-3 carrier through the restriction enzyme digestion cleft cutting, make up bacterium fusion expression vector pGEX-4T-3-mbFGF;
(2) with fusion expression vector pGEX-4T-3-mbFGF transformed into escherichia coli BL21 (DE3) cell, the cell growth conditions is 200 rev/mins, 37 ℃ of temperature;
(3) as thalli growth OD
595When value is 0.5-0.8, add the inductor IPTG of 1.0mM, induced 2-3 hour at 25 ℃.In the time of 4 ℃, with 5,000 * g centrifugal 5 minutes, collect thalline;
(4) thalline is after PBS buffer solution for cleaning 3 times, ultrasonication.Smudge cells with 15,000 * g centrifugal 1 hour, is collected supernatant in the time of 4 ℃;
(5) supernatant is cycled through Glutathione Sepharose 4B affinity chromatographic column, 4 ℃ are spent the night.1 * PBS flushing of 10 times of bed volumes of not conjugated protein usefulness;
(6) bonded GST-mbFGF fusion rotein is cut and use the elutriant wash-out with zymoplasm at post, collect eluate;
(7) above-mentioned eluate is adsorbed zymoplasm through Benzamidine Sepharose 4B Fast Flow affinity chromatographic column, behind wash-out, obtain proteantigen mbFGF.
3, with immunize rabbit behind the fused antigen of freund's adjuvant, obtain the anti-mbFGF polyclonal antibody of rabbit through separation and purification, concrete preparation process is following:
(1) with the mbFGF proteantigen with after freund's adjuvant mixes, fully emulsified.Adopt back multi-point injection method immunizing rabbit.The about 100 μ g of antigen amount of each immunity, immunity three times.Each immune two weeks are surveyed antibody titer after arteria auricularis is got blood examination.
(2) with the antigen coated enzyme plate of 500 μ g/ml concentration, 100 μ l/ holes, 4 ℃ are spent the night;
(3) washing after, add serum sample to be checked, 100 μ l/ holes, 37 ℃ 1 hour; If negative control hole;
(4) after the washing, add the antibody reagent of the goat anti-rabbit igg of HRP mark, 100 μ l/ holes, 37 ℃ of lucifuges developed the color 20 minutes; Use 2.5M H
2SO
4After the termination reaction, read the A490 value in each hole.
To the mbFGF antibody of preparation, detect through the ELISA method, antibody capable combines with the mbFGF specificity, and its relation conefficient reaches level of signification, tires＞1: 6000, and the result is as shown in Figure 7.
4, the anti-mbFGF polyclonal antibody of rabbit obtains detecting the immune latex microsphere of recombined small-mouse Prostatropin through carboxyl-amino chemical coupling method coupling activation microballoon
After the mbFGF Antibody Preparation was accomplished, through chemical coupling monodisperse carboxyl polystyrene microsphere, preparation was used to detect the proteic immune latex microsphere of mbFGF, and concrete preparation and detection step are following:
(1) with the monodisperse carboxyl polystyrene microsphere with acvator EDC (carbodiimide) activation, add protective material Sulfo-NHS then and make microballoon form stable active ester midbody;
(2) add anti-mbFGF polyclonal antibody, incubation 1-2 hour;
(3) after the washing, collect immune microsphere;
(4) through ELIASA, adopt enzyme-linked immunosorbent assay (ELISA) method, detect mbFGF sample (as shown in Figure 8).
Through the ELISA method validation, the immune latex microsphere LDL that is used to detect mbFGF is recombined small-mouse Prostatropin 60pg/ml, coefficient R
2=0.96, the result is as shown in Figure 9.This shows that the present invention is prepared, and to be used to detect the immune latex microsphere detection sensitivity of mbFGF higher, and simple to operate, repeatability is strong, has shown good prospects for application.
Claims (7)
1. detect the immune latex microsphere of recombined small-mouse Prostatropin; It is characterized in that it being to use the monodisperse carboxyl polystyrene microsphere to be substrate material; Immune latex microsphere with the anti-mbFGF polyclonal antibody acquisition of carboxyl-amino chemical coupling method coupling; Be the microballoon that encapsulates by anti-mbFGF protein polyclone antibody, microsphere particle size 50nm.
2. the preparation method of the immune latex microsphere of the described detection recombined small-mouse of claim 1 Prostatropin is characterized in that concrete steps are following:
(1) modifies and makes up total length mouse bFGF expressed sequence again according to the preferential codon of procaryotic cell expression;
(2) mouse bFGF gene fragment is gone in the glutathione-S-transferase gst gene fusion expression vector through the restriction enzyme digestion cleft cutting, make up the bacterium fusion expression vector;
Said glutathione-s-transferase gene fusion expression vector is pGEX-4T-3, and the fusion expression vector that makes up after accomplishing is pGEX-4T-3-mbFGF;
(3) the bacterium fusion expression vector is carried out bacterial expression after, utilize the affinity chromatography column purification to obtain the GST-mbFGF fusion rotein; Said affinity chromatographic column is Glutathione Sepharose 4B;
(4) utilize zymoplasm at post cutting GST-mbFGF fusion rotein, eluate obtains mouse bFGF after affinity chromatographic column is removed zymoplasm; Affinity chromatographic column is Benzamidine Sepharose 4B Fast Flow:
(5) with immunize rabbit behind the fused antigen of freund's adjuvant, obtain the anti-mbFGF polyclonal antibody of rabbit through separation and purification; Said antigen is mbFGF albumen;
(6),, obtain detecting the immune latex microsphere of recombined small-mouse Prostatropin through carboxyl-amino chemical coupling method coupling activation microballoon with the anti-mbFGF polyclonal antibody of the resulting rabbit of step (5);
Said microballoon is the monodisperse carboxyl polystyrene microsphere, big or small 50nm, and it is carboxyl that group is carried on the surface,
The activation of said microballoon is at first to use acvator carbodiimide activation microballoon, adds protective material Sulfo-NHS then and makes microballoon form stable active ester midbody, in order to connecting antibody molecule.
3. the preparation method of the immune latex microsphere of detection recombined small-mouse Prostatropin according to claim 2 is characterized in that the concrete steps of modifying and make up total length mouse bFGF expressed sequence according to the preferential codon of procaryotic cell expression again are:
(1) design total length mbFGF gene order is modified according to the preferential codon of procaryotic cell expression again, adopts full gene synthesis technology to make up total length mbFGF gene, and totally 462 pairs of bases are cloned into carrier pGEM-T Easy, and the result is as shown in Figure 2.
(2) exactness of inserting through bacterium colony PCR checking mbFGF gene segment, the result is as shown in Figure 3.
(3) will clone mbFGF gene bacterium and implement amplification cultivation, and extract and carry out that enzyme is cut and agarose electrophoretic analysis behind the plasmid, the result is as shown in Figure 4.
(5) dna sequencing guarantees that the full sequence 100% of gene is accurate
4. the preparation method of the immune latex microsphere of detection recombined small-mouse Prostatropin according to claim 2; It is characterized in that mouse bFGF gene fragment is gone among the glutathione-S-transferase gst gene fusion expression vector pGEX-4T-3 through the restriction enzyme digestion cleft cutting, the concrete operations step that makes up bacterium fusion expression vector pGEX-4T-3-mbFGF is following:
(1) the mbFGF gene fragment is gone in the pGEX-4T-3 carrier through the restriction enzyme digestion cleft cutting, make up bacterium fusion expression vector pGEX-4T-3-mbFGF;
(2) with fusion expression vector pGEX-4T-3-mbFGF transformed into escherichia coli BL21 (DE3) cell, the cell growth conditions is 200 rev/mins, 37 ℃ of temperature;
(3) as thalli growth OD
595When value is 0.5-0.8, add the inductor IPTG of 1.0mM, induced 2-3 hour at 25 ℃.In the time of 4 ℃, with 5,000 * g centrifugal 5 minutes, collect thalline;
(4) thalline is after PBS buffer solution for cleaning 3 times, ultrasonication.Smudge cells with 15,000 * g centrifugal 1 hour, is collected supernatant in the time of 4 ℃;
(5) supernatant is cycled through Glutathione Sepharose 4B affinity chromatographic column, 4 ℃ are spent the night.1 * PBS flushing of 10 times of bed volumes of not conjugated protein usefulness;
(6) bonded GST-mbFGF fusion rotein is cut and use the elutriant wash-out with zymoplasm at post, collect eluate;
(7) above-mentioned eluate is adsorbed zymoplasm through Benzamidine Sepharose 4B Fast Flow affinity chromatographic column, behind wash-out, obtain proteantigen mbFGF.
5. the preparation method of the immune latex microsphere of detection recombined small-mouse Prostatropin according to claim 2; It is characterized in that the concrete preparation process that obtains the anti-mbFGF polyclonal antibody of rabbit through separation and purification is following with immunize rabbit behind the fused antigen of freund's adjuvant:
(1) with the mbFGF proteantigen with after freund's adjuvant mixes, fully emulsified.Adopt back multi-point injection method immunizing rabbit.The about 100 μ g of antigen amount of each immunity, immunity three times.Each immune two weeks are surveyed antibody titer after arteria auricularis is got blood examination.
(2) with the antigen coated enzyme plate of 500 μ g/ml concentration, 100 μ l/ holes, 4 ℃ are spent the night;
(3) washing after, add serum sample to be checked, 100 μ l/ holes, 37 ℃ 1 hour; If negative control hole;
(4) after the washing, add the antibody reagent of the goat anti-rabbit igg of HRP mark, 100 μ l/ holes, 37 ℃ of lucifuges developed the color 20 minutes; Use 2.5M H
2SO
4After the termination reaction, read the A490 value in each hole.
6. the preparation method of the immune latex microsphere of detection recombined small-mouse Prostatropin according to claim 2; It is characterized in that the anti-mbFGF polyclonal antibody of rabbit through carboxyl-amino chemical coupling method coupling activation microballoon, the concrete preparation process of immune latex microsphere that obtains detecting the recombined small-mouse Prostatropin is following:
(1) with the monodisperse carboxyl polystyrene microsphere with acvator EDC (carbodiimide) activation, add protective material Sulfo-NHS then and make microballoon form stable active ester midbody;
(2) add anti-mbFGF polyclonal antibody, incubation 1-2 hour;
(3) after the washing, collect the immune latex microsphere that detects the recombined small-mouse Prostatropin.
7. the application of the immune latex microsphere of the said described detection recombined small-mouse Prostatropin of claim 1 is characterized in that through ELIASA, adopts enzyme-linked immunosorbent assay, detects the bFGF of mbFGF sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103395428A CN102432685A (en) | 2011-11-01 | 2011-11-01 | Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103395428A CN102432685A (en) | 2011-11-01 | 2011-11-01 | Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102432685A true CN102432685A (en) | 2012-05-02 |
Family
ID=45981069
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103395428A Pending CN102432685A (en) | 2011-11-01 | 2011-11-01 | Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102432685A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104474533A (en) * | 2014-11-24 | 2015-04-01 | 华南理工大学 | Method for preparing compound microspheres for repairing wound |
| CN105699660A (en) * | 2016-02-24 | 2016-06-22 | 浙江理工大学 | Specific immunity detection method of ancient wool |
| CN108192905A (en) * | 2018-01-25 | 2018-06-22 | 南华大学 | A kind of expression cassette, expression vector, the method for host strain and preparation without label recombined human IL37 mature peptides |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050130234A1 (en) * | 1996-12-20 | 2005-06-16 | Oystein Fodstad | Method for characterization of abnormal cells |
| CN101871935A (en) * | 2010-06-07 | 2010-10-27 | 吕军 | Covalent cross-linking method for carboxylic polystyrene microsphere and C reaction protein antibody |
| CN102109521A (en) * | 2010-12-16 | 2011-06-29 | 中国环境科学研究院 | Immunolatex microsphere for detecting CpTI and preparation method thereof |
-
2011
- 2011-11-01 CN CN2011103395428A patent/CN102432685A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050130234A1 (en) * | 1996-12-20 | 2005-06-16 | Oystein Fodstad | Method for characterization of abnormal cells |
| CN101871935A (en) * | 2010-06-07 | 2010-10-27 | 吕军 | Covalent cross-linking method for carboxylic polystyrene microsphere and C reaction protein antibody |
| CN102109521A (en) * | 2010-12-16 | 2011-06-29 | 中国环境科学研究院 | Immunolatex microsphere for detecting CpTI and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张惠菊等: "液态芯片技术在临床实验室诊断中的应用", 《医学研究生学报》, vol. 22, no. 4, 30 April 2009 (2009-04-30), pages 422 - 426 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104474533A (en) * | 2014-11-24 | 2015-04-01 | 华南理工大学 | Method for preparing compound microspheres for repairing wound |
| CN105699660A (en) * | 2016-02-24 | 2016-06-22 | 浙江理工大学 | Specific immunity detection method of ancient wool |
| CN108192905A (en) * | 2018-01-25 | 2018-06-22 | 南华大学 | A kind of expression cassette, expression vector, the method for host strain and preparation without label recombined human IL37 mature peptides |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103543261B (en) | Cattle and sheep Brucellosis indirect enzyme-linked immunosorbent assay antibody assay kit and preparation method thereof | |
| CN103059109B (en) | Mycoplasma pneumonia antigen, preparation method and immunodetection kit | |
| CA2833081C (en) | Hypoallergen | |
| CN102432685A (en) | Immune latex microspheres for detecting recombined mouse basic fibroblast growth factor and preparation method and application of immune latex microspheres | |
| CN103969234A (en) | Method capable of detecting four kinds of type I diabetes autoimmune antibodies simultaneously | |
| CN106636004B (en) | TMV-CMV-PVY triple virus colloidal gold rapid detection test strip | |
| CN104610443B (en) | A kind of high stability restructuring Procalcitonin, Preparation method and use | |
| CN105541977B (en) | Riemerella anatipestifer OmpH intercept recombinant protein and preparation method and application | |
| CN102998457B (en) | The Competitive assays enzyme-linked immune detection method of grass carp TNFa lpha | |
| CN111518174B (en) | Optimized African swine fever CD2v protein and high-efficiency expression method and application thereof | |
| CN105541992A (en) | Human MBP 18.5kD variant antigen epitope peptide and specific detection kits | |
| CN102585010A (en) | Immunolatex microsphere for detecting rhbFGF (Recombinant Human Basic Fibroblast Growth Factor) and preparation method thereof | |
| CN114773460B (en) | Nano antibody of targeted rotavirus protein and application thereof | |
| CN102109521A (en) | Immunolatex microsphere for detecting CpTI and preparation method thereof | |
| CN103074302A (en) | Anti-C-peptide monoclonal antibody and use thereof in detection of C-peptide content | |
| CN102775486A (en) | Novel reagent and kit for renal injury monitoring | |
| JP2017520246A5 (en) | ||
| CN112730829B (en) | Colloidal gold test strip for rapidly detecting oyster herpesvirus OsHV-1 and application thereof | |
| CN102643332B (en) | Application of B cell epitope peptide of human PCT (Procalcitonin) and monoclonal antibody of B cell epitope peptide | |
| Yu et al. | A conformational change of C fragment of tetanus neurotoxin reduces its ganglioside-binding activity but does not destroy its immunogenicity | |
| CN110540599B (en) | Klebsiella pneumoniae Elisa detection kit based on Klebsiella pneumoniae surface protein antibody and preparation method thereof | |
| CN102391375A (en) | Method for producing antibody by coupling multi-polypeptide epitope of protein antigen with carrier | |
| CN102323421A (en) | Enzyme linked immunosorbent assay kit and preparation method thereof | |
| JP2007527523A (en) | Reagent, method and kit for detecting feed enzymes | |
| WO2022158373A1 (en) | Protein, hemolytic streptococcus vaccine, dna, vector, transformant, method for producing protein, baculovirus, agrobacterium, latex particle, kit, and method for measuring anti-streptolysin o antibody |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120502 |