CN114316041B - Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card - Google Patents
Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card Download PDFInfo
- Publication number
- CN114316041B CN114316041B CN202111639563.1A CN202111639563A CN114316041B CN 114316041 B CN114316041 B CN 114316041B CN 202111639563 A CN202111639563 A CN 202111639563A CN 114316041 B CN114316041 B CN 114316041B
- Authority
- CN
- China
- Prior art keywords
- nls
- monoclonal antibody
- rarα
- immunization
- antibody
- 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.)
- Active
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a preparation method of an NLS-RAR alpha monoclonal antibody, a rapid detection card based on the antibody and a preparation method thereof. The preparation method of the monoclonal antibody mainly comprises the following steps: determining immunogen of target protein, performing experimental animal immunization, fusing the immunized animal spleen cells with myeloma cells to obtain hybridoma cells, screening positive clones and preparing a large amount of monoclonal antibodies. And then preparing an APL rapid detection card based on the NLS-RARalpha specific monoclonal antibody. The monoclonal antibody prepared by the invention has good specificity; APL rapid detection card based on NLS-RAR alpha monoclonal antibody has the characteristics of being minimally invasive, convenient, rapid and economical, can be used for detecting peripheral blood, does not need special equipment, is an advantage which the existing diagnosis and treatment technology does not have, and is easier to popularize than the existing technology.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a preparation method of an NLS-RAR alpha monoclonal antibody, a rapid detection card based on the antibody and a preparation method thereof.
Background
Acute promyelocytic leukemia (Acute Promyelocytic Leukemia, APL) belongs to the M3 subtype in FAB typing, a special type of acute leukemia with specific chromosomal changes and fusion genes. APL frequently develops severe hemorrhage and Disseminated Intravascular Coagulation (DIC) in early stages, and is always considered as the most serious acute leukemia, and although the targeting induction effect of all-trans retinoic acid and arsenical agents is reliable, 15% of APL patients still have early death due to untimely diagnosis.
Currently, APL diagnosis relies mainly on bone marrow morphology, chromosome detection, fluorescence In Situ Hybridization (FISH), immunotyping and fusion gene detection techniques. Bone marrow morphology is a direct meat eyeglass to examine bone marrow cell morphology for typing diagnosis, and even experienced persons often confuse with other types of leukemia due to cell morphology heterogeneity and polymorphism, which brings clinical trouble and even misdiagnosis. Chromosome detection and Fluorescence In Situ Hybridization (FISH) can detect chromosomal karyotype changes of APL canonical t (15; 17) (q 22; q 21); the immunophenotyping is to detect APL cell surface immune markers CD33, CD13 and the like by using flow cytometry; and the fusion gene detection is to detect the characteristic leukemia-retinoic acid receptor alpha (PML-RARalpha) fusion gene by using real-time quantitative PCR (RQ-PCR). These techniques are more objective and sensitive than morphology, but there are also limitations: (1) expensive equipment (flow cytometry, immunofluorescence microscope, etc.); (2) the detection period is long (days are needed); (3) the technical requirements of detection personnel are high; (4) the economic cost is high, and the burden of patients is heavy; (5) the bone marrow sample is needed, peripheral blood cannot be detected, materials are not easy to obtain, and the wound of a patient is large. Thus, it is highly desirable to explore new technologies that can detect APL early, quickly, and objectively.
APL is characterized genetically by the t (15; 17) (q 22; q 21) chromosomal ectopic and PML-RARα fusion genes, the latter encoding the PML-RARα fusion proteins formed are key to the development of APL. Most of the previous researches take PML-RARα fusion proteins as a whole, however, the previous innovations of the team find that NLS-RARα proteins generated by cracking PML-RARα fusion proteins by NE are another key index for promoting the occurrence and development of APL and are novel diagnostic markers for early diagnosis of APL.
Disclosure of Invention
In order to solve the technical problems, the invention provides a preparation method of an NLS-RAR alpha monoclonal antibody and an APL rapid detection card based on the antibody, wherein the detection card is based on an NLS-RAR alpha novel protein marker as a core, and the rapid detection card for APL is developed by preparing the NLS-RAR alpha specific monoclonal antibody and combining with an immune related technology.
The technical scheme of the invention is as follows:
the invention provides a preparation method of an NLS-RARalpha monoclonal antibody, which comprises the following steps:
s1, selecting and designing an immunogen according to an NLS-RAR alpha target protein sequence;
s2, performing experimental animal immunization by using the immunogen designed by the S1;
s3, fusing spleen cells of the experimental animal subjected to S2 immunization with myeloma cells to obtain hybridoma cells;
s4, culturing the hybridoma cells fused in the S3 for 2-3 weeks, taking culture solution supernatant for ELISA detection, and primarily screening positive clones;
s5, verifying and screening again by adopting an immunoblotting method to obtain positive clones with good specificity and high antibody concentration;
s6, preparing a large amount of monoclonal antibodies.
As a preferred embodiment of the invention, the immunogen is COM-1a, the sequence of the COM-1a is VQSVPGAHPVPV-C, the corresponding antibody of the COM-1a is 1a, and the 1a polypeptide is directed against the NE cleavage site of PML-RARA, so that the NLS-RARA specific polypeptide can be specifically identified.
As a preferred embodiment of the invention, the animal selected for immunization of the animal in S2 is a healthy New Zealand white rabbit.
As a preferred embodiment of the invention, the specific steps of the immunization of the experimental animal in S2 are as follows: selecting healthy New Zealand white rabbits to carry out animal immunization according to a conventional experimental rabbit injection immunization method, wherein the total injection time is 5 needles, the injection immunization interval is 3 weeks, 2 weeks and 2 weeks, and the COM-1a immunogen needs to be mixed with a proper adjuvant 1:1 before the immunization injection, so that the full emulsification is carried out to ensure the effect of the immunization injection. As a preferred embodiment of the invention, the polypeptides used in ELISA detection in S4 are COM-1a and COM-1b, the sequence of the COM-1b is VPGAHPVPVYA-C, the corresponding antibodies of the COM-1b are 1b and 1b polypeptides, and the polypeptides are not specific and can recognize NLS-RAR alpha and PML-RAR alpha simultaneously.
As a preferred embodiment of the present invention, the criteria for preliminary screening of positive clones in S4 are: clone culture supernatant reacts with CQM-1a-BSA and CQM-1b-BSA proteins simultaneously, and the clone with double positive property is regarded as NLS-RARalpha and PML-RARalpha; clones that reacted positively only with the CQM-1a-BSA protein are considered to be more likely to recognize only NLS-RARα specific cleavage fragments, i.e., the clones of interest for selection.
As a preferred embodiment of the present invention, the criteria for screening positive clones in S5 are: the supernatant of the clone culture solution is respectively subjected to immunoblotting verification with the protein extracts of 293T cells transfected with NLS-RAR alpha plasmid and 293T cells transfected with empty control plasmid, and can react with the 293T cell protein extract transfected with NLS-RAR alpha plasmid but not react with the 293T cell protein extract transfected with empty, so that the clone is ideal clone.
As a preferred embodiment of the present invention, the specific steps of S6 are: and (3) preparing an antibody mRNA template by cell lysis of the positive clone selected in the S5, carrying out RT-PCR reaction on the template to obtain heavy chain cDNA and light chain cDNA of the antibody, carrying out sequencing confirmation, constructing an expression vector, preparing an NLS-RAR alpha recombinant monoclonal antibody by transfecting cells with the constructed antibody expression plasmid, and purifying the antibody by a protein purification column to obtain the monoclonal antibody.
The invention provides a preparation method of an APL rapid detection card based on an NLS-RARalpha monoclonal antibody, which comprises the following steps:
s1, labeling an NLS-RARalpha specific monoclonal antibody by using immune colloidal gold;
s2, re-dissolving the precipitate by using a gold re-dissolving solution, and then spreading the precipitate on a gold pad with the thickness of 6mm multiplied by 150mm for later use;
s3, coating the NLS-RARalpha specific monoclonal antibody stock solution and the diluent on an NC membrane detection line and a quality control line respectively, and drying at low humidity for later use;
s4, selecting a bottom plate, sequentially adhering water absorbing paper, an NC film, a gold pad and a sample chromatographic pad from top to bottom, and assembling to obtain the rapid detection reaction plate for NLS-RARalpha.
The invention provides an APL rapid detection card based on an NLS-RARalpha monoclonal antibody, which is prepared by adopting the preparation method.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention provides a preparation method of an NLS-RAR alpha monoclonal antibody, which designs an immunogen with good specificity according to an NLS-RAR alpha target protein sequence, thereby obtaining the NLS-RAR alpha monoclonal antibody with good specificity.
2. The APL rapid detection card based on the NLS-RAR alpha monoclonal antibody has the characteristics of being minimally invasive, convenient, rapid and economical, can be used for detecting peripheral blood, does not need special equipment, is an advantage which is not possessed by the existing diagnosis and treatment technology, and is easier to popularize than the existing technology.
Drawings
FIG. 1 shows NLS-RARα target protein sequence;
FIG. 2 is an ELISA validation of NLS-RARα animal immune serum;
FIG. 3 shows WB validation and screening of positive clones;
FIG. 4 is a verification of three clones # 36, #47, # 49;
FIG. 5 shows CQM-1a-BSA# 36 monoclonal antibody titer assay;
FIG. 6 shows SDS-PAGE electrophoresis to verify NLS-RARα recombinant monoclonal antibody specificity;
FIG. 7 is a schematic diagram of the APL rapid detection technique;
fig. 8 shows the result interpretation rule of APL rapid detection technology.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings.
The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
Example 1
Preparation of NLS-RARα monoclonal antibodies
NLS-RARα target protein sequences are shown in FIG. 1, and immunogens are selected and designed based on analysis of structural properties, fragment immunogenicity, and water solubility of NLS-RARα sequences.
The polypeptide immunogen sequence was determined and is shown in Table 1. Wherein the COM-1a sequence VQSVPGAHPVPV-C is 1-12 amino acid sites of NLS-RAR alpha, and the corresponding 1a polypeptide can specifically recognize NLS-RARA specific polypeptide aiming at NE cleavage sites of PML-RAR alpha. The COM-1b sequence VPGAHPVPVYA-C is 4-14 amino acid locus of NLS-RAR alpha, and the corresponding 1b polypeptide has no specificity, can theoretically recognize NLS-RAR alpha and PML-RAR alpha, and is mainly used for screening and contrast verification. After the sequence is determined, the immunogen is synthesized by artificial chemistry. The synthesized polypeptide is subjected to molecular weight analysis by LC-MS, the accuracy of an amino acid sequence is determined, the purity is detected by using an HPLC method, the purity is ensured to be not lower than 90%, and the polypeptide is used as an immunogen for experimental animal immunization after meeting the requirements.
TABLE 1 immunogen sequences for NLS-RARA monoclonal antibody preparation
| Peptide fragment | Name of the name | Sequence(s) | Use of the same |
| Pepitde1 | COM-1a | VQSVPGAHPVPV-C | For immunization |
| Pepitde2 | COM-1b | VPGAHPVPVYA-C | For screening and control verification |
Immunization of experimental animals was performed with the synthesized polypeptide immunogen COM-1 a:
selecting healthy New Zealand white rabbits to carry out animal immunization according to a conventional experimental rabbit injection immunization method, wherein the total injection time is 4-5 needles, the injection immunization interval is 3+2+2+2 (weeks), and the immunogen needs to be mixed with a proper adjuvant 1:1 before the immunization injection, so that the effect of the immunization injection is ensured. During the period, a small amount of rabbit serum is collected and subjected to titer detection by using an ELISA method, and the immune effect is evaluated. The results show that: serum from both rabbits immunized (numbers 1141, 1142) was able to react with the immunogen COM-1a and U937 cell (NR) protein extracts transfected with NLS-rarα plasmid, in accordance with expectations. The results are shown in FIG. 2, where NC represents the U937 protein extract of the transfected empty control plasmid, NR represents the U937 cell protein extract of the transfected NLS-RARα plasmid (with HA tag), and COM1a represents the COM-1a immunogen. (1) pre-1141: no. 1141 rabbit preimmune serum; (2) pre-1142: serial No. 1142 rabbit preimmune serum; (3) anti-1141: serum 1 month after immunization of the number 1141 rabbit; (4) anti-1142: serum 1 month after immunization of the number 1142 rabbit; (5) anti-HA: anti-HA antibodies were used as experimental controls.
Spleen cells of the experimental animal are isolated after the immunization is finished, and the spleen cells and myeloma cells are fused to develop hybridoma cells. After supplementing nutrition such as FBS, the cell mixture subjected to the fusion operation is kept stand in a cell culture box for 2-3 weeks, hybridoma culture supernatants are taken for ELISA detection, positive hybridoma clones are screened in a primary mode, and OD (sups 1:20) > 1.0 is regarded as positive clones. Among them, CQM-1a-BSA and CQM-1b-BSA double-positive clones demonstrated that NLS-RARα and PML-RARα could be recognized simultaneously. Only CQM-1a-BSA positive clones demonstrated a greater likelihood of recognizing only NLS-RARA specific fragments, and the results are shown in Table 2.
Table 2. Immunization of elisa screening of hybridoma clones
To further screen positive clones with higher antibody titers, the specificity and sensitivity of the antibodies are ensured. The empty vector (NC) and NLS-RARalpha (NR) plasmids are firstly introduced into 293T cells, and meanwhile, the immunoblotting (WB) method is adopted to verify and screen clone bodies again, so that the results show that: three clones # 36, #47, #49 were better in specificity and higher in antibody concentration, and the results are shown in FIGS. 3 and 4, wherein (1) 293T-NC: 293T cells transfected with empty control plasmid (2.) (293T-NR: 293T cells transfected with NLS-RARα plasmid by lentivirus.
CQM-1a-BSA # 36 positive clone is selected, an antibody mRNA template is prepared after cell lysis, the template is combined with a heavy chain gene and a light chain gene (cDNA) of an antibody are amplified by using a specially designed primer for RT-PCR reaction, and the heavy chain gene and the light chain gene are established to an expression vector after sequencing and confirmation. The constructed antibody expression plasmid is transfected into HEK293 cells to prepare NLS-RAR alpha recombinant monoclonal antibodies, after the antibodies are purified by a protein purification column, the concentration of the monoclonal antibodies is detected by ELISA, see figure 5, and the specificity of the NLS-RAR alpha recombinant monoclonal antibodies is again verified by SDS-PAGE electrophoresis, see figure 6. The purified antibodies were stored in PBS buffer at low temperature.
Example 2
APL rapid detection card based on NLS-RAR alpha monoclonal antibody
The immune colloidal gold is used for marking NLS-RARalpha specific monoclonal antibody, and a gold complexing solution is used for complexing the precipitate and then is paved on a gold pad with the thickness of 6mm multiplied by 150mm for drying for standby. And respectively coating the NLS-RAR alpha antibody stock solution and the diluent on a detection line and a quality control line on the NC film, and drying at low humidity for later use. And selecting a bottom plate, sequentially adhering water absorbing paper, an NC film, a gold pad and a sample chromatographic pad from top to bottom, and assembling to obtain the rapid detection reaction plate for NLS-RARalpha. The schematic diagram of the rapid detection technology is shown in fig. 7, and the result judgment rule is shown in fig. 8.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (7)
1. A method for preparing an NLS-RARα monoclonal antibody, which is characterized by comprising the following steps:
s1, selecting and designing an immunogen according to an NLS-RAR alpha target protein sequence, wherein the immunogen is COM-1a, the sequence of the COM-1a is VQSVPGAHPVPV-C, wherein the C end represents the carboxyl end in the sequence direction, the antibody corresponding to the COM-1a is 1a, the COM-1a polypeptide immunogen is designed aiming at the NE cleavage site of PML-RAR alpha, and specific antibodies for specifically recognizing NLS-RAR alpha are generated by immunization;
s2, performing experimental animal immunization by using the immunogen designed by the S1;
s3, fusing spleen cells of the experimental animal subjected to S2 immunization with myeloma cells to obtain hybridoma cells;
s4, culturing the hybridoma cells fused in the S3 for 2-3 weeks, taking culture solution supernatant for ELISA detection, and primarily screening positive clones;
s5, verifying and screening again by adopting an immunoblotting method to obtain positive clones with good specificity and high antibody concentration;
s6, preparing a large amount of monoclonal antibodies.
2. The method of claim 1, wherein the animals selected for immunization of S2 are healthy new zealand white rabbits.
3. The method for preparing the NLS-RARα monoclonal antibody according to claim 1, wherein the specific step of immunization of the experimental animal in S2 is as follows: selecting healthy New Zealand white rabbits to carry out animal immunization according to a conventional experimental rabbit injection immunization method, wherein the total injection time is 5 needles, the injection immunization interval is 3 weeks, 2 weeks and 2 weeks, and the COM-1a immunogen needs to be mixed with an adjuvant 1:1 before the immunization injection, so that the full emulsification is carried out to ensure the effect of the immunization injection.
4. The method for preparing the NLS-RARα monoclonal antibody according to claim 1, wherein polypeptides used in ELISA detection in S4 comprise COM-1a and COM-1b, the sequence of the COM-1b is VPGAHPVPVYA-C, wherein the C end represents the carboxyl end in the sequence direction, the corresponding antibody of the COM-1b is 1b,1b is not specific, the COM-1b immunogen is designed aiming at the common sequence of NLS-RARα and PML-RARα, and the specific antibodies generated by immunization can simultaneously recognize NLS-RARα and PML-RARα.
5. The method for preparing the NLS-RARα monoclonal antibody according to claim 1, wherein the criteria for preliminary screening of positive clones in S4 are: clone culture supernatant reacted with CQM-1a-BSA and CQM-1b-BSA proteins simultaneously, and double positive clone was regarded as identifying NLS-RARα and PML-RARα simultaneously; only clones that reacted positively with the CQM-1a-BSA protein are considered to recognize only NLS-RARα specific cleavage fragments, i.e., the clones of interest for screening.
6. The method for preparing the NLS-RARα monoclonal antibody according to claim 1, wherein the criteria for screening positive clones in S5 are: performing immunoblotting verification on clone culture fluid supernatant and protein extracts of 293T cells transfected with NLS-RAR alpha plasmid and 293T cells transfected with empty control plasmid respectively, and reacting with the 293T cell protein extract transfected with NLS-RAR alpha plasmid but not with the 293T cell protein extract transfected with empty to obtain ideal clone.
7. The method for preparing the NLS-RARα monoclonal antibody according to claim 1, wherein the specific steps of S6 are as follows: and (3) preparing an antibody mRNA template by cell lysis of the positive clone selected in the S5, carrying out RT-PCR reaction on the template to obtain heavy chain cDNA and light chain cDNA of the antibody, carrying out sequencing confirmation, constructing an expression vector, preparing an NLS-RAR alpha recombinant monoclonal antibody by transfecting cells with the constructed antibody expression plasmid, and purifying the antibody by a protein purification column to obtain the monoclonal antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111639563.1A CN114316041B (en) | 2021-12-29 | 2021-12-29 | Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111639563.1A CN114316041B (en) | 2021-12-29 | 2021-12-29 | Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114316041A CN114316041A (en) | 2022-04-12 |
| CN114316041B true CN114316041B (en) | 2023-06-13 |
Family
ID=81016324
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111639563.1A Active CN114316041B (en) | 2021-12-29 | 2021-12-29 | Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114316041B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974071A (en) * | 2010-09-26 | 2011-02-16 | 中国人民解放军第三军医大学 | Insulin B chain HLA-A*0201 restrictive CTL epitope modified peptide ligand and acquisition method and application thereof |
| WO2011062997A2 (en) * | 2009-11-17 | 2011-05-26 | Musc Foundation For Research Development | Human monoclonal antibodies to human nucleolin |
| CN103399157A (en) * | 2013-07-30 | 2013-11-20 | 重庆医科大学附属永川医院 | Locating method of PML (promyelocytic leukemia) protein with deletion of nuclear localization signal |
| CN104031149A (en) * | 2014-06-16 | 2014-09-10 | 重庆医科大学附属永川医院 | Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis |
| CN109929848A (en) * | 2019-04-03 | 2019-06-25 | 西北农林科技大学 | A kind of human tumor suppressor gene PML saltant type and its application |
-
2021
- 2021-12-29 CN CN202111639563.1A patent/CN114316041B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011062997A2 (en) * | 2009-11-17 | 2011-05-26 | Musc Foundation For Research Development | Human monoclonal antibodies to human nucleolin |
| CN101974071A (en) * | 2010-09-26 | 2011-02-16 | 中国人民解放军第三军医大学 | Insulin B chain HLA-A*0201 restrictive CTL epitope modified peptide ligand and acquisition method and application thereof |
| CN103399157A (en) * | 2013-07-30 | 2013-11-20 | 重庆医科大学附属永川医院 | Locating method of PML (promyelocytic leukemia) protein with deletion of nuclear localization signal |
| CN104031149A (en) * | 2014-06-16 | 2014-09-10 | 重庆医科大学附属永川医院 | Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis |
| CN109929848A (en) * | 2019-04-03 | 2019-06-25 | 西北农林科技大学 | A kind of human tumor suppressor gene PML saltant type and its application |
Non-Patent Citations (2)
| Title |
|---|
| NLS-RARα对APL细胞分化的影响及其机制研究;李健;中国学术期刊（光盘版）;第2020卷(第12期);1-64 * |
| Requirement of the coiled-coil domain of PML-RARα oncoprotein for localization, sumoylation, and inhibition of monocyte differentiation;Young-Eui Kim ET.AL;Requirement of the coiled-coil domain of PML-RARα oncoprotein for localization, sumoylation, and inhibition of monocyte differentiation;第330卷(第3期);746-754 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114316041A (en) | 2022-04-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8420348B2 (en) | Antibodies for the detection of integrin complexes in FFPE material | |
| CN103998467B (en) | Anti-folacin receptor Alpha antibodies and application thereof | |
| US8492520B2 (en) | Binding epitopes for G250 antibody | |
| EP2900265B1 (en) | Anti-uroplakin ii antibodies systems and methods | |
| CN107108733B (en) | Anti-active GIP antibody | |
| CN111925992B (en) | Hybridoma cell strain secreting anti-Wnt-7 a monoclonal antibody, monoclonal antibody and application thereof | |
| CN114316041B (en) | Preparation method of NLS-RAR alpha monoclonal antibody, rapid detection card based on antibody and preparation method of rapid detection card | |
| US7666603B2 (en) | Breast cancer related protein, gene encoding the same, and method of diagnosing breast cancer using the protein and gene | |
| CN112646036B (en) | anti-CD 23 specific monoclonal antibody and application thereof | |
| CN115925866A (en) | Monoclonal antibody for breast cancer detection and kit thereof | |
| CN112540176B (en) | Kit, method and computer-readable storage medium for diagnosing diseases associated with FAP expression abnormality | |
| CN112646037B (en) | anti-CD 105 specific monoclonal antibody and application thereof | |
| JPWO2009044561A1 (en) | Anti-proNT / NMN monoclonal antibody | |
| KR102235935B1 (en) | Monoclonal antibody N16-F2 specific to human naive pluripotent stem cells | |
| CN110878122B (en) | Recombinant anti-PD-L1 monoclonal antibodies | |
| CN102131829A (en) | Antibodies against human epo receptor | |
| CN105949312B (en) | Anti-human PCT antibody and its application | |
| CN106046157B (en) | Human PCT fluorescent quantitative detection test paper card | |
| CN112250767B (en) | Antibody combined with Strep-Tag II label and application thereof | |
| CN111705039B (en) | Hybridoma cell strain secreting anti-SOX 17 monoclonal antibody and application | |
| JP4187249B2 (en) | Presynaptic protein p120 | |
| WO2022241752A1 (en) | ANTIBODY OR ANTIBODY FRAGMENT THAT SPECIFICALLY BINDS TO VOLTAGE-GATED SODIUM CHANNEL α SUBUNIT NAV1.7 | |
| Marzouq et al. | Production of Recombinant Human EGFR CR2 Domain and Generation of Monoclonal Antibodies Discriminating The R and K Variants. | |
| CN116410322A (en) | Monoclonal antibody against human PCDH7 protein and application thereof | |
| WO2023154780A1 (en) | Anti-her2/neu antibodies and methods of use |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |