WO2020155208A1 - Nano-anticorps, adsorbant utilisant ledit nano-anticorps comme ligand et utilisation associée - Google Patents

Nano-anticorps, adsorbant utilisant ledit nano-anticorps comme ligand et utilisation associée Download PDF

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WO2020155208A1
WO2020155208A1 PCT/CN2019/075439 CN2019075439W WO2020155208A1 WO 2020155208 A1 WO2020155208 A1 WO 2020155208A1 CN 2019075439 W CN2019075439 W CN 2019075439W WO 2020155208 A1 WO2020155208 A1 WO 2020155208A1
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nanobody
adsorbent
amino acid
acid sequence
tnf
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王玉凤
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康元医疗科技(大连)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
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C07K16/241—Tumor Necrosis Factors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • 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
    • C07K2317/50—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
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®

Definitions

  • the invention belongs to the field of biotechnology, and relates to a nanobody-based immunoaffinity material, in particular to a nanobody, an adsorbent using the nanobody as a ligand, and uses thereof.
  • the adsorbent includes a carrier matrix and a nanobody as a ligand.
  • the material uses a nanobody that specifically recognizes, for example, TNF- ⁇ as a ligand, and can reduce the excessive TNF- ⁇ level in the patient's body through blood purification, thereby alleviating and/ Or treat diseases such as sepsis.
  • Tumor necrosis factor is a cytokine that can induce necrosis of human tumor cells. It is a trimer structure composed of three identical polypeptide chains, which can be combined with TNF receptor (TNF-R) to mediate a variety of biological processes. Among them, TNF-R exists on the surface of a variety of normal and tumor cells. The mechanism of downstream signal transmission after binding to TNF is unclear. It may be related to the activation of protein kinase C (PKC), which catalyzes the phosphorylation of receptor proteins.
  • PPC protein kinase C
  • TNF can be divided into two types: TNF- ⁇ and TNF- ⁇ .
  • TNF- ⁇ is mainly derived from monocytes and macrophages, and the molecular weight of each subunit of its homotrimer is 17kDa.
  • TNF- ⁇ As an inflammatory mediator, TNF- ⁇ has a dual effect on the human body: under normal circumstances, TNF- ⁇ presents a protective response to the body, and can participate in the process of resisting infection by bacteria, viruses and parasites, promote tissue repair, and cause Tumor cell apoptosis, through the inflammatory response to clear the body's pathogenic factors, plays an important regulatory role in maintaining the stability of the internal environment and the renewal of tissues.
  • TNF- ⁇ presents a protective response to the body, and can participate in the process of resisting infection by bacteria, viruses and parasites, promote tissue repair, and cause Tumor cell apoptosis, through the inflammatory response to clear the body's pathogenic factors, plays an important regulatory role in maintaining the stability of the internal environment and the renewal of tissues.
  • certain diseases such as sepsis and rheumatoid arthritis
  • the excessively high blood concentration can lead to an intensified inflammatory response and cause irreversible damage to the patient's body tissues.
  • TNF- ⁇ enhances the activation and differentiation of macrophages during the onset of sepsis in animals and humans, and releases other pro-inflammatory factors (such as IL-6, IL-8 and MIF), lipid molecules, and responsiveness. Oxygen free radicals and nitrogen free radicals amplify the inflammatory cascade and induce organ dysfunction.
  • TNF- ⁇ can increase the body's secretion and synthesis of chemokines and the expression of cell adhesion molecules, thereby increasing the recruitment rate of lymphocytes and monocytes.
  • TNF- ⁇ can also stimulate the expression of antigen-expressing myosin heavy chain (MHC) type II receptors, and induce synovial fibroblasts and chondrocytes to produce prostaglandin E2 (PGE2) and matrix metalloproteinase (MMP) (collagenase) And matrix protease), leading to inflammation and hyperplasia of synovium, and destruction of cartilage tissue (see Non-Patent Document 1).
  • MHC myosin heavy chain
  • MMP matrix metalloproteinase
  • TNF- ⁇ has also been found to be closely related to the pathogenesis of a variety of malignant tumors, inflammatory bowel disease (IBD), type 2 diabetes (T2D), Crohn’s disease, immunosuppressive disease and multiple sclerosis disease . Therefore, reducing the excess TNF- ⁇ in the blood circulation of patients or neutralizing its activity through different ways is of great significance for the prevention, alleviation, treatment and reduction of complications of related diseases.
  • IBD inflammatory bowel disease
  • T2D type 2 diabetes
  • Crohn’s disease immunosuppressive disease
  • multiple sclerosis disease multiple sclerosis disease
  • TNF- ⁇ There are two main ways to treat TNF- ⁇ : one is to inhibit the activity of TNF- ⁇ or TNF- ⁇ receptors in the body through antibody drugs to block TNF- ⁇ from working; the other is to use blood purification
  • the method directly adsorbs and removes TNF- ⁇ in the blood of patients.
  • Intestinal syndrome inflammatory bowel syndrome
  • multiple sclerosis multiple sclerosis
  • Addison's disease Addison's disease
  • autoimmune hepatitis autoimmune hepatitis
  • treatment methods based on monoclonal antibody drugs often have the following problems in clinical use: (1) Although the blood level of TNF- ⁇ in the patient is very low, in order to achieve the necessary blood concentration to obtain the drug effect, a greater need For example, the single dose of tocilizumab is 8mg/kg, and each treatment requires an injection of about 400mg, which makes it difficult to avoid high treatment costs. For example, AbbVie’s adalimumab sells as high as 195 Yuan/mg, the average annual treatment cost is 202,000 yuan. (2) High-dose antibody drug infusion also has a higher safety risk.
  • Non-Patent Document 2 In clinical trials of anti-TNF- ⁇ monoclonal antibodies, patients are prone to a variety of adverse reactions, which can cause severe bacterial infections, affect nerve function, and increase the risk of cancer in the lymphatic system and immune diseases (see Non-Patent Document 2).
  • Blood purification is another effective means to interfere with blood components, which can be used to remove toxic or pathogenic substances from the blood.
  • Commonly used clinical blood purification techniques include hemoperfusion, hemofiltration, hemodialysis, etc., and have been successfully applied to the treatment of renal failure, liver failure, drug poisoning and autoimmune diseases in clinical practice.
  • blood purification treatment methods and medical materials for inflammatory factors have also been rapidly developed, and have been used in the treatment of sepsis, rheumatoid arthritis, multiple organ failure, severe pancreatitis and other diseases.
  • CytoSorbTM adsorbent (CytoSorbents Corporation, Monmouth Junction, NJ) is a typical resin-based hydrophobic adsorbent, which has been reported to be used for the adsorption and removal of inflammatory factors.
  • the adsorption medium uses polystyrene-divinylbenzene copolymer particles. When applied to hemoperfusion, the total clearance rate of most cytokines in the blood can reach more than 80% within 120 minutes. However, whether CytoSorbTM also filters the critical blood components, nutrients and drugs in the patient's blood during the treatment, and the safety risks caused by this still need to be further evaluated. Zhuhai Jianfan Biotechnology Co., Ltd.
  • Nankai University invented a nano-composite microsphere adsorbent that can remove inflammatory factors in patients.
  • IA immunoadsorption
  • ligands adsorption functional groups
  • IA immunosorbents
  • IA has higher selectivity, safety and specificity for eliminating endogenous or exogenous pathogenic factors in the blood.
  • Nanobodies have the advantages of good water solubility, acid and alkali resistance, high temperature resistance, high stability, easy expression, weak immunogenicity, high sensitivity, and good tissue penetration. In addition, various test results prove , Various modifications to the Nanobody will not damage its structural integrity, can be compatible with a high-throughput screening platform, and have the advantages of low cost and convenient preparation.
  • Patent Literature 5 discloses an aflatoxin M1 nanobody 2014AFM-G2, and successfully uses silica gel microspheres or agarose gel as a solid phase carrier to prepare immunosorbents and immunoaffinity adsorption columns for agricultural products and The purification stage of aflatoxin samples in food.
  • Patent Document 6 discloses a highly specific Nanobody against c-Myc, which can purify recombinant proteins containing c-Myc tags. The Capture Select series of chromatographic separation media under Thermo Fisher uses nanobodies as ligands to purify antibodies, antibody fragments and proteins through affinity solutions.
  • Dalian University of Technology applied nanobody-based immunoadsorbent materials to the field of blood purification, and developed a specific blood purification adsorbent for ⁇ 2 microglobulin in patients with renal failure (see Patent Document 7).
  • the highly selective TNF- ⁇ immunoadsorbent based on Nanobody of the present invention will further expand the application of Nanobody in the field of blood purification treatment.
  • Patent Document 1 International Publication WO2004/041862
  • Patent Document 2 International Publication WO2006/122786
  • Patent Document 3 CN103585977B
  • Patent Document 4 CN106334541A
  • Patent Document 5 CN103869065A
  • Patent Document 6 CN106890622A
  • Patent Document 7 CN104098694B
  • Non-Patent Document 1 Kalden JR. Emerging role of anti-tumor necrosis factor therapy in rheumatic diseases. Arthritis Res, 2002, 4 (Suppl 2): S34-S40.
  • Non-Patent Literature 2 Hu Xue, Liu Yang. Research progress and adverse reactions of tumor necrosis factor- ⁇ inhibitors. Hebei Medicine, 2012, 11, 34: 3477-3480.
  • Non-Patent Document 3 Long Haibo, Zhang Xun, Hou Fanfan, Experimental study on the treatment of endotoxin shock with immunoadsorbent anti-medium. Chinese Journal of Trauma, 1998, 14(2): 92-95
  • Non-Patent Document 4 Wang Qin, Wu Xiongfei, Wang Junxia, etc., the development of specific tumor necrosis factor- ⁇ immunoadsorbent column and the experimental study of its adsorption rate. Journal of the Third Military Medical University, 2003, 25(24): 2220-2222
  • the present invention is based on solving the existing technical problems, and its purpose is to provide a Nanobody, an adsorbent using the Nanobody as a ligand, and the aforementioned Nanobody or the aforementioned adsorbent for preparing reagents for removing TNF- ⁇ And/or use in medical devices, and these uses in immunoassay, enrichment and/or purification.
  • the first aspect of the present invention is a first aspect of the present invention.
  • the amino acid sequence of the aforementioned Nanobody includes a complementarity determining region and a framework region.
  • the aforementioned complementarity determining region includes complementarity determining region 1 (CDR1), complementarity determining region 2 (CDR2) and complementarity determining region 3 ( CDR3),
  • CDR1 complementarity determining region 1
  • CDR2 complementarity determining region 2
  • CDR3 complementarity determining region 3
  • the amino acid sequence of the aforementioned framework region 1 is SEQ ID NO: 4,
  • the amino acid sequence of the aforementioned framework region 2 is SEQ ID NO: 5,
  • the amino acid sequence of the aforementioned framework region 3 is SEQ ID NO: 6,
  • the amino acid sequence of the aforementioned framework region 4 is SEQ ID NO: 7.
  • Nanobody according to 1 or 2 wherein the Nanobody further includes a hinge region, and the amino acid sequence of the hinge region is SEQ ID NO: 8.
  • Nanobody according to 1 or 2 wherein the Nanobody is produced by introducing a gene into Escherichia coli or yeast and using Escherichia coli or yeast expression.
  • the second aspect of the present invention is a first aspect of the present invention.
  • An adsorbent is provided, the adsorbent includes a carrier matrix and the Nanobody described in 1 to 4 above.
  • the carrier matrix is a porous material
  • the porous material is selected from one of agarose gel microspheres, cellulose beads, magnetic beads, silica gel microspheres, activated carbon, and resin microspheres .
  • the third aspect of the present invention is a first aspect of the present invention.
  • Nanobody described in 1 to 4 or the adsorbent described in 5 to 8 in immunoassay, enrichment and/or purification.
  • the adsorbent of the present invention has a nanobody with a special structure and a carrier matrix, the nanobody can specifically recognize and bind to TNF- ⁇ , thereby effectively removing bad inflammatory mediators from the blood, and because the nanobody itself has low immunogenicity and safety It has the advantages of high sex, so the adsorbent is used in the field of blood purification, which is very safe and effective, and helps to relieve and treat kidney failure, sepsis, rheumatoid arthritis and other diseases.
  • the adsorbent of the present invention has a large adsorption capacity
  • the nanobody itself also has the properties of acid and alkali resistance, high salt resistance, high temperature resistance, etc., so that the immunoaffinity adsorbent can be applied to the enrichment and purification of TNF- ⁇ .
  • FIG. 1 is the electrophoresis diagram of TNF- ⁇ Nanobody expression in whole bacteria
  • FIG. 1 is an electrophoresis diagram of TNF- ⁇ Nanobody after purification.
  • ligand means that in affinity chromatography, biomolecules that are coupled to a carrier and used to separate specific binding substances are often called ligands.
  • solid loading refers to the total amount of ligands coupled per unit volume of the affinity medium (adsorbent).
  • the amino acid sequence of the Nanobody comprises a complementarity determining region and a framework region.
  • the aforementioned complementarity determining region includes complementarity determining region 1 (CDR1), complementarity determining region 2 (CDR2) and complementarity determining region 3 (CDR3), the aforementioned
  • the amino acid sequence of the complementarity determining region 1 (CDR1) is SEQ ID NO:1
  • the amino acid sequence of the aforementioned complementarity determining region 2 (CDR2) is SEQ ID NO: 2
  • the amino acid sequence of the aforementioned complementarity determining region 3 (CDR3) is SEQ ID NO :3.
  • the complementarity determining regions of different Nanobodies are quite different. As long as the complementarity determining regions in the Nanobodies satisfy the above-mentioned amino acid sequence, the effects of the present invention can be achieved.
  • an amino acid sequence with a sequence homology of 50% or more is preferable, an amino acid sequence with a sequence homology of 70% or more is more preferable, and an amino acid sequence with a sequence homology of 95% or more is most preferable.
  • the framework region of the aforementioned Nanobody includes framework region 1 (FR1), framework region 2 (FR2), framework region 3 (FR3) and framework region 4 (FR4), and the amino acid sequence of the aforementioned framework region 1 (FR1) is SEQ ID NO: 4, the amino acid sequence of the aforementioned framework region 2 (FR2) is SEQ ID NO: 5, the amino acid sequence of the aforementioned framework region 3 (FR3) is SEQ ID NO: 6, the amino acid sequence of the aforementioned framework region 4 (FR4) It is SEQ ID NO: 7, but the amino acid sequence of these framework regions of the present invention is not limited to this, and can be appropriately selected according to the matching with the complementarity determining region.
  • the Nanobody further includes a hinge region, and the amino acid sequence of the hinge region is preferably SEQ ID NO: 8, but the amino acid sequence is not limited thereto.
  • the hinge region By including the hinge region, the flexibility of the Nanobody is increased.
  • Nanobodies are produced by introducing genes into Escherichia coli or yeast and using Escherichia coli or yeast expression.
  • the Nanobody of the present invention as the ligand of the carrier matrix, can be prepared by methods known in the art. As long as the Nanobody of the present invention can be obtained, it can be prepared, for example, by the following method: First, construct the nanobody Non-immune library, and obtain Nanobody gene through screening technology. Secondly, introduce Nanobody gene into Escherichia coli or yeast by means of genetic engineering recombination to obtain genetically engineered bacteria for expressing Nanobody, and then use this genetic engineering The bacteria induce the expression of the nanobody, and finally it is purified by metal chelation chromatography to obtain the pure nanobody that meets the requirements.
  • the carrier matrix of the present invention is preferably a porous material, and the porous material is preferably selected from one or more of agarose gel microspheres, cellulose beads, magnetic beads, silica gel microspheres, activated carbon, and resin microspheres.
  • Carriers for the aforementioned adsorbents can be purchased commercially, as specific examples of products, such as Sepharose CL-6B (GE Healthcare, US), Nanomicro series of resin microspheres (Suzhou Nanomicro Technology Co., Ltd.), but It is not limited to these products.
  • the activation can be, for example, but not limited to the following methods: firstly, epoxy activation, secondly diaminopropylimine (DADPA) activation, and finally iodoacetic acid activation, etc.
  • the aforementioned adsorbent is obtained by coupling the Nanobody to an activated carrier.
  • the specific method is not particularly limited.
  • the purified Nanobody can be reduced to obtain a Nanobody solution, and then the Nanobody solution can be combined with the carrier. Mix, centrifuge, and finally clean and filter the gel to obtain the final adsorbent.
  • the adsorbent of the present invention can preferably be used to specifically recognize TNF- ⁇ .
  • the nanobody or adsorbent of the present invention can be used to prepare reagents for removing TNF- ⁇ , and can also be used to prepare medical devices for removing TNF- ⁇ .
  • the Nanobody or adsorbent of the present invention can also be used in immunodetection, enrichment and/or purification applications.
  • the phage display library used in the present invention is a non-immune library with T7 phage as a carrier.
  • the establishment steps are as follows:
  • the first round of nested PCR uses cDNA as the template, and the upstream primer 1 and downstream primer 1 are respectively the upstream and downstream primers. After amplification, a band with a size of 650-750bp is recovered and used as the template for the second round of PCR , The upstream and downstream primers are the upstream primer 2 and the downstream primer 2, respectively. PCR products of 450 ⁇ 500bp are recovered;
  • Upstream primer 1 5’-GGTACGTGCT GTTGAACTGT TCC-3’
  • Downstream primer 1 5’-CTTGGTGGTCCTGGCTGCTCT-3’
  • Upstream primer 2 5’-AAGCTTTTGT GGTTTTGGTG TCTTGGGTTC-3’
  • Downstream primer 2 5’-AAGCTTGGGG TCTTCGCTGT GGTGCG-3’
  • T7 vector with T4 ligase ( 10-3Cloning Kit, Merck Millipore 70550-3) and VHH gene fragments;
  • the titer of the original library is 1.5 ⁇ 1010 pfu/mL, and the diversity is 2.0 ⁇ 107.
  • the antigen was diluted with TBS to 10 ⁇ g/mL, 100 ⁇ L was added to a 96-well plate, and incubated at 4°C for 12h.
  • Aspirate the antigen diluent in the well wash the plate 3 times with TBS, pat dry, add 1% protein-free blocking solution (purchased from Shenggong Bioengineering Co., Ltd.), 300 ⁇ L/well, incubate at room temperature for 2h (1% protein-free during screening Blocking fluid and 1% BSA are used alternately).
  • Aspirate the blocking agent from the well wash the plate 6 times with TBST, pat dry, add the amplified phage, 100 ⁇ L/well, and incubate at room temperature for 30 min. Wash the plate 10 times with TBST, add T7 elution buffer (1% SDS) to elute the phage, incubate at room temperature for 30 min, and amplify the eluate for the next round of screening.
  • the screening eluate is subjected to solid amplification, plaques are picked, and the plaque amplification fluid is used as a template, and UPprimer3 and DOWNprimer3 are used as the upstream and downstream primers for PCR amplification. increase;
  • Upstream primer 3 5’-GGAGCTGTCGTATTCCAGTC-3’
  • Downstream primer 3 5'-AACCCCTCAAGACCCGTTTA-3';
  • amino acid sequence of CDR1 is SEQ ID NO: 1
  • amino acid sequence of CDR2 is SEQ ID NO: 2
  • amino acid sequence of CDR3 is SEQ ID NO: 3
  • FR1 The amino acid sequence of FR2 is SEQ ID NO: 4
  • amino acid sequence of FR2 is SEQ ID NO: 5
  • amino acid sequence of FR3 is SEQ ID NO: 6
  • amino acid sequence of FR4 is SEQ ID NO: 7.
  • the strain was inoculated into a shake flask at an inoculum of 1%, and cultured overnight at 37°C with 170 r/min shaking.
  • the activated seeds were inoculated into a large culture flask according to the inoculum of 1% and continued to be cultured.
  • IPTG IPTG with a final concentration of 0.2 mM was added for overnight induction.
  • centrifuge at 3900r/min for 20 minutes to obtain wet bacteria containing TNF- ⁇ Nanobody. SDS-PAGE was performed on the whole bacteria, and the electrophoresis results are shown in Figure 1, where the loading volume of 2 was 20 ⁇ L, and the loading volume of 3 was 30 ⁇ L.
  • the chromatographic eluate is subjected to SDS-PAGE to determine the purity.
  • the electrophoresis result is shown in Figure 2.
  • the purified protein has a purity of more than 90%, which can be used for subsequent adsorbent synthesis and adsorbent evaluation.
  • the agarose gel is thoroughly washed with 1mol/L NaCl solution and water in sequence, and it can be stored in 20% ethanol at 4°C.
  • the experimental method refers to the 2015 edition of "The Pharmacopoeia of the People's Republic of China” and the “Biological Evaluation of Medical Devices” Chapter 11 "Selection of Interaction with Blood”
  • n (A1-A2)*100%/(A3-A2), where n is the hemolysis rate (%), A1 is the absorbance of the test product group, and A2 is the negative control group Absorbance, A3 is the absorbance of the positive control group.
  • the adsorbent with a solid load of 18.0 mg/mL has a TNF- ⁇ adsorption capacity of 19.43 mg/mL, and the corresponding adsorption molar ratio is 0.98, close to 1, the adsorption rate at this time is 75.8%.
  • TNF- ⁇ kit (Solarbio, SEKH-0047) to measure the concentration before and after adsorption to determine the adsorption in the blood.
  • the removal rate of TNF- ⁇ in the serum of the adsorbent can reach 55.2%, which can reduce the blood TNF- ⁇ level of simulated liver cancer patients to the level of healthy people (the blood TNF- ⁇ level of healthy people is 0.39 ⁇ 0.16ng/mL, refer to "Plasma Tumor Necrosis Factor-Content Changes in Patients with Acute Cerebral Infarction"), it can be seen that the adsorbent of the present invention is of great significance in the field of blood purification treatment.
  • the nanobody of the present invention as well as the adsorbent with a special structure of the nanobody and a carrier matrix, can specifically recognize and bind to TNF- ⁇ , thereby effectively removing bad inflammatory mediators from the blood, and because the nanobody itself has low immunogenicity and safety It has the advantages of high sex, so the adsorbent is used in the field of blood purification, which is very safe and effective, and helps to relieve and treat kidney failure, sepsis, rheumatoid arthritis and other diseases.
  • the adsorbent of the present invention has a large adsorption capacity, and the nanobody itself has the properties of acid and alkali resistance, high salt resistance, high temperature resistance, etc., so that the immunoaffinity adsorbent can be applied to the enrichment and purification of TNF- ⁇ .

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Abstract

L'invention concerne un nano-anticorps, un adsorbant utilisant ledit nano-anticorps comme ligand et une utilisation associée. La séquence d'acides aminés du nano-anticorps comprend des régions déterminant la complémentarité et des régions cadre, dans les régions déterminant la complémentarité, la séquence d'acides aminés de la région déterminant la complémentarité 1 (CDR1) étant SEQ ID NO : 1 ; la séquence d'acides aminés de la région déterminant la complémentarité 2 (CDR2) étant SEQ ID NO : 2 ; et la séquence d'acides aminés de la région déterminant la complémentarité 3 (CDR3) étant SEQ ID NO : 3. Le nano-anticorps selon l'invention et l'adsorbant l'utilisant comme ligand reconnaissent de manière spécifique des antigènes, ce qui permet d'éliminer efficacement des médiateurs inflammatoires indésirables dans le sang et d'obtenir un bon effet de purification du sang.
PCT/CN2019/075439 2019-01-29 2019-02-19 Nano-anticorps, adsorbant utilisant ledit nano-anticorps comme ligand et utilisation associée WO2020155208A1 (fr)

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