WO2023245801A1 - Anticorps monoclonal anti-candida mannane et son utilisation - Google Patents

Anticorps monoclonal anti-candida mannane et son utilisation Download PDF

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WO2023245801A1
WO2023245801A1 PCT/CN2022/108079 CN2022108079W WO2023245801A1 WO 2023245801 A1 WO2023245801 A1 WO 2023245801A1 CN 2022108079 W CN2022108079 W CN 2022108079W WO 2023245801 A1 WO2023245801 A1 WO 2023245801A1
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monoclonal antibody
candida
seq
variable region
mannan
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Chinese (zh)
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付成华
魏新宇
翟栓柱
盛长忠
粟艳
周泽奇
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丹娜(天津)生物科技股份有限公司
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/14—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from fungi, algea or lichens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0681—Cells of the genital tract; Non-germinal cells from gonads
    • C12N5/0682—Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0684—Cells of the urinary tract or kidneys
    • C12N5/0686—Kidney cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56961—Plant cells or fungi
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/106—Plasmid DNA for vertebrates
    • C12N2800/107—Plasmid DNA for vertebrates for mammalian
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/37—Assays involving biological materials from specific organisms or of a specific nature from fungi
    • G01N2333/39—Assays involving biological materials from specific organisms or of a specific nature from fungi from yeasts
    • G01N2333/40—Assays involving biological materials from specific organisms or of a specific nature from fungi from yeasts from Candida
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/10—Detection of antigens from microorganism in sample from host
    • 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

Definitions

  • the invention belongs to the field of biotechnology, and specifically relates to an anti-Candida mannan monoclonal antibody and its application.
  • Candida albicans is a clinically isolated opportunistic human pathogenic fungus. Especially in patients with downregulated immunity, such as organ transplant patients, HIV-infected patients, etc., it can cause a wide range of superficial and Deep systemic infections include the oral cavity, female vagina, etc., causing thrush, vaginitis, etc. It can also invade epidermis and endothelial cells, enter the bloodstream, and reach internal organs, such as kidneys, brain, etc., causing sepsis, which can even lead to death in severe cases.
  • the gold standard for detecting Candida infection is blood culture.
  • the disease progresses rapidly.
  • Traditional culture methods have low positive detection and long culture cycles, which often lead to missed diagnosis and misdiagnosis, leading to delayed disease progression. Therefore, it is of great significance to develop a highly specific antibody that can be used for Candida detection.
  • Monoclonal antibodies are highly uniform antibodies produced by a single B cell clone and only target a specific antigenic epitope. They are called monoclonal antibodies. Usually prepared using hybridoma technology. Hybridoma antibody technology is based on cell fusion technology, which fuses sensitized B cells with the ability to secrete specific antibodies and myeloma cells with unlimited reproduction ability into B cell hybrids. tumor. By cultivating a single hybridoma cell with this characteristic into a cell population, a specific antibody, that is, a monoclonal antibody, can be prepared against an antigenic epitope.
  • CN105866409A discloses a Candida mannan antigen immunodetection kit, which includes a Mn-coated enzyme-labeled plate and an anti-Mn polyclonal enzyme-labeled antibody.
  • the kit coats Mn on an enzyme plate, competes the sample to be tested or standard antigen with the coated antigen for binding to limited antibody binding sites, and then reacts with the substrate through horseradish peroxidase for color development. , draw a standard curve based on the Mn standard substance, and calculate the concentration value of the antigen to be detected based on the standard curve.
  • the test kit has good sensitivity and specificity, is simple and easy to operate, and detects quickly and sensitively. But because polyclonal antibodies detect a range of epitopes on a protein, they are subject to a higher risk of batch-to-batch variation than monoclonal antibodies.
  • Immunoassay technology is a method that utilizes the specific binding reaction between antigens and antibodies to achieve qualitative or quantitative detection of antigens or antibodies by detecting markers labeled on the reactants. According to the different labeling substances, it is divided into enzyme-linked immunoassay technology (Enzyme-Linked Immunosorbant Assay, ELISA), immunofluorescence detection technology, chemiluminescence immunoassay technology, immune microsphere technology, immunocolloidal gold technology, etc. Among them, chemiluminescence immunoassay technology has the characteristics of simple operation, high sensitivity and short detection time, and is widely used in clinical detection and scientific research.
  • enzyme-linked immunoassay technology Enzyme-Linked Immunosorbant Assay, ELISA
  • immunofluorescence detection technology chemiluminescence immunoassay technology
  • immune microsphere technology immune microsphere technology
  • immunocolloidal gold technology etc.
  • chemiluminescence immunoassay technology has the characteristics of simple operation, high sensitivity and short detection time, and is widely used
  • Antibodies with excellent performance are the basis of immunoassay technology. Only with good antibodies can immunoassay kits with excellent performance be developed. Therefore, providing an anti-Candida mannan monoclonal antibody that recognizes more sites, has stronger affinity and specificity is of great significance in Candida detection.
  • the purpose of the present invention is to provide an anti-Candida mannan monoclonal antibody and its application.
  • the anti-Candida mannan monoclonal antibody is a rabbit-derived monoclonal antibody, which has the characteristics of multiple antigen recognition sites, good specificity, and high affinity. It solves the practical application problems of mouse-derived monoclonal antibodies and provides Candida Provides new solutions for bacterial detection, diagnosis, prevention and treatment.
  • the present invention provides an anti-Candida mannan monoclonal antibody, which includes a heavy chain variable region and a light chain variable region;
  • the light chain variable region also includes the light chain CDR1 shown in SEQ ID NO.4 and the light chain CDR2 shown in SEQ ID NO.5.
  • the nucleotide sequence encoding the heavy chain variable region of the anti-Candida mannan monoclonal antibody includes the sequence shown in SEQ ID NO. 9, and the nucleotide sequence encoding the anti-Candida mannan monoclonal antibody.
  • the nucleotide sequence of the light chain variable region includes the sequence shown in SEQ ID NO. 10.
  • the host cells include 293F cells or CHO cells.
  • the present invention provides a pharmaceutical composition comprising the anti-Candida mannan monoclonal antibody described in the first aspect.
  • the present invention provides a kit for detecting Candida, which includes the anti-Candida mannan monoclonal antibody described in the first aspect.
  • the kit for detecting Candida also includes any one or a combination of at least two of a positive control substance, a negative control substance, an antibody diluent, a chromogenic solution, a stop solution, a blocking solution or a washing solution.
  • the present invention provides the anti-Candida mannan monoclonal antibody described in the first aspect, the nucleic acid molecule described in the second aspect, the expression vector described in the third aspect, the host cell described in the fourth aspect, or The use of any one or a combination of at least two of the pharmaceutical compositions described in the fifth aspect in the preparation of candida infection treatment drugs and/or detection products.
  • the present invention has the following beneficial effects:
  • the monoclonal antibody in the present invention has many original recognition sites, good specificity and high affinity; the monoclonal antibody is a rabbit-derived monoclonal antibody and has many antigen recognition sites; the monoclonal antibody does not interact with other sugars (including Capsular polysaccharide, galactomannan, lipopolysaccharide, peptidoglycan, 1,3- ⁇ -D-glucan and BSA) cross-react with strong specificity; the screened monoclonal antibodies have strong resistance to Mn antigen The binding ability and affinity are high, and the titer for Mn antigen reaches 1:1280000 (OD value>0.5).
  • Figure 2 is the detection result of the affinity activity of the monoclonal antibody in Example 7.
  • FIG. 3 shows the cross-reaction results in Example 8.
  • the reconstituted solution is subjected to alcohol precipitation, and the precipitate is collected.
  • the precipitate is the crude Candida mannan (Mn), and the precipitate is reconstituted with deionized water.
  • dialysis, and the dialysis product is filtered through a 0.22 ⁇ m filter membrane, and the obtained product is the crude Candida mannan product.
  • the fifth week Boost immunity.
  • the immunization method is the same as the third week.
  • the immunization dose is 1mL per animal.
  • Week 6 The first serum titer test. Take 200-500 ⁇ L of ear marginal venous blood from the immunized New Zealand big-eared rabbits, let it stand for 30 minutes, then centrifuge to separate the serum, and measure the antiserum titer.
  • Week 11 Strengthen immunity, the immunization method is the same as step (3).
  • the immunized New Zealand big-eared rabbits were sacrificed, and the spleens were removed under aseptic conditions. Wash once with cell culture medium, grind, pass through a stainless steel mesh, wash twice with cell culture medium, and centrifuge to obtain cells.
  • the obtained hybridoma cells were screened by ELISA method to find up to 10 wells of polyclonal antibody cell lines targeting Candida mannan antigen.
  • the cell lines can produce specific monoclonal antibodies with high affinity against the respective antigens.
  • the cell fusion step there are two parental cells and three randomly fused cells in the culture medium.
  • the successfully fused hybridoma cells must be separated from the numerous cells.
  • B lymphocytes cannot survive in vitro for a long time. Only myeloma cells and their own fused cells need to be removed. Therefore, the fused cells need to be cultured in HAT culture medium to selectively retain hybridoma cells.
  • the growth of the cells can be observed on the 5th day after fusion.
  • the indirect ELISA method can be used to detect the cell culture supernatant and screen positive hybridoma cell lines for cloning culture.
  • the positive hybridoma cells were cloned and cultured using the limiting dilution method.
  • the positive hybridoma cells with the strongest detection result titer were expanded until the cell positivity rate reached 100% to determine the strain.
  • Use ELISA to measure the potency of the culture supernatant of the established hybridoma cell line, and freeze the expanded cultured monoclonal hybridoma cell line in liquid nitrogen.
  • RNA is then precipitated from the aqueous layer using isopropyl alcohol.
  • DNA was precipitated from the organic layer using ethanol.
  • Proteins were precipitated from the phenol-ethanol supernatant using isopropanol precipitation. Wash the precipitated RNA to remove impurities and resuspend it for later use.
  • RNA as the template
  • tRNA as the primer
  • a cDNA single strand complementary to the RNA template is synthesized, which forms an RNA-cDNA hybrid with the RNA template. body.
  • the RNA strand is hydrolyzed, and the second DNA strand is synthesized using cDNA as a template. At this point, total RNA was reverse transcribed into cDNA.
  • Antibody fragments of heavy and light chains were amplified according to the standard operating procedure of Rapid Amplification of cDNA Ends (RACE). The amplified antibody fragments were individually cloned into standard cloning vectors. Perform colony PCR to screen for clones with the correct size insert. The antibody sequence is obtained.
  • RACE Rapid Amplification of cDNA Ends
  • the nucleotide sequence encoding the heavy chain variable region of the anti-Candida mannan monoclonal antibody includes the sequence shown in SEQ ID NO. 9; the light chain encoding the anti-Candida mannan monoclonal antibody may The nucleotide sequence of the variable region includes the sequence shown in SEQ ID NO.10.
  • Cell culture medium harvest Centrifuge to collect the cell culture medium supernatant, and use ProteinABeads for antibody purification;
  • Example 7 Using ELISA method to detect the affinity activity (titer) of monoclonal antibodies against Candida mannan (Mn)
  • Example 8 Using ELISA method to detect the specificity of monoclonal antibodies
  • the specificity of monoclonal antibodies is detected through cross-reaction.
  • the steps of cross-reaction are as follows:
  • the enzyme plate was coated with mannan, galactomannan, capsular polysaccharide, lipopolysaccharide, peptidoglycan, 1,3- ⁇ -D-glucan and BSA respectively, and the coating amount per well was 50ng/ hole; dilute the monoclonal antibody to 10ng/mL, add it to each microplate, add 100 ⁇ L to each well, and incubate at 37°C for 1 hour; after washing, add 100 ⁇ L of secondary antibody (HRP-labeled goat anti-rabbit IgG) to each well, and incubate at 37°C. Incubate for 0.5h; add TMB after washing, incubate at 37°C for 15min, and terminate reading.
  • the cross-reaction results are shown in Figure 3. The results show that the obtained monoclonal antibody does not cross-react with other sugars and has strong specificity.
  • the present invention provides an anti-Candida mannan monoclonal antibody.
  • the monoclonal antibody is a rabbit-derived monoclonal antibody with multiple antigen recognition sites, good specificity, and high affinity. It solves the problem of mouse-derived monoclonal antibodies. There are problems in the practical application of antibodies.
  • the anti-Candida mannan monoclonal antibody has important application value in preparing products for Candida detection.

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Abstract

La présente demande concerne un anticorps monoclonal anti-candida mannane et son utilisation. L'anticorps monoclonal est un anticorps monoclonal dérivé de lapin, a de multiples sites de reconnaissance d'antigène, une bonne spécificité et une affinité élevée, et a une valeur d'application importante dans la préparation de produits de détection de candida.
PCT/CN2022/108079 2022-06-24 2022-07-27 Anticorps monoclonal anti-candida mannane et son utilisation WO2023245801A1 (fr)

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CN202210728744.XA CN114933652B (zh) 2022-06-24 2022-06-24 一种抗念珠菌甘露聚糖单克隆抗体及其应用
CN202210728744.X 2022-06-24

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Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989003398A1 (fr) * 1987-10-14 1989-04-20 Teijin Limited Anticorps monoclonal humain contre le candida
WO2000052053A1 (fr) * 1999-03-01 2000-09-08 Research And Development Institute, Inc. Anticorps diriges contre le phosphomannane et protecteurs contre les candidoses
CN103204929A (zh) * 2012-01-16 2013-07-17 天津贻诺琦生物工程有限公司 一种白色念珠菌多克隆抗体的制备方法
WO2014174293A1 (fr) * 2013-04-25 2014-10-30 The University Court Of The University Of Aberdeen Molécules d'anticorps antifongiques et leurs utilisations
CN105866409A (zh) * 2016-06-27 2016-08-17 丹娜（天津）生物科技有限公司 一种念珠菌甘露聚糖抗原免疫检测试剂盒及其制备方法与应用
CN105968198A (zh) * 2016-06-27 2016-09-28 天津汇滨生物科技有限公司 一种念珠菌甘露聚糖的单克隆抗体及其制备方法
WO2016172660A1 (fr) * 2015-04-23 2016-10-27 Board Of Regents Of The Nevada System Of Higher Education, On Behalf Of The University Of Nevada, Reno Détection fongique à l'aide d'épitope de mannane
CN106117353A (zh) * 2016-06-27 2016-11-16 丹娜（天津）生物科技有限公司 一种念珠菌甘露聚糖Mn抗原的多克隆抗体及其制备方法
CN110885372A (zh) * 2019-12-17 2020-03-17 丹娜（天津）生物科技有限公司 一种抗念珠菌甘露聚糖的兔源单克隆抗体及其应用

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989003398A1 (fr) * 1987-10-14 1989-04-20 Teijin Limited Anticorps monoclonal humain contre le candida
WO2000052053A1 (fr) * 1999-03-01 2000-09-08 Research And Development Institute, Inc. Anticorps diriges contre le phosphomannane et protecteurs contre les candidoses
CN103204929A (zh) * 2012-01-16 2013-07-17 天津贻诺琦生物工程有限公司 一种白色念珠菌多克隆抗体的制备方法
WO2014174293A1 (fr) * 2013-04-25 2014-10-30 The University Court Of The University Of Aberdeen Molécules d'anticorps antifongiques et leurs utilisations
WO2016172660A1 (fr) * 2015-04-23 2016-10-27 Board Of Regents Of The Nevada System Of Higher Education, On Behalf Of The University Of Nevada, Reno Détection fongique à l'aide d'épitope de mannane
CN105866409A (zh) * 2016-06-27 2016-08-17 丹娜（天津）生物科技有限公司 一种念珠菌甘露聚糖抗原免疫检测试剂盒及其制备方法与应用
CN105968198A (zh) * 2016-06-27 2016-09-28 天津汇滨生物科技有限公司 一种念珠菌甘露聚糖的单克隆抗体及其制备方法
CN106117353A (zh) * 2016-06-27 2016-11-16 丹娜（天津）生物科技有限公司 一种念珠菌甘露聚糖Mn抗原的多克隆抗体及其制备方法
CN110885372A (zh) * 2019-12-17 2020-03-17 丹娜（天津）生物科技有限公司 一种抗念珠菌甘露聚糖的兔源单克隆抗体及其应用

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