CN114426584A - 一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 - Google Patents
一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 Download PDFInfo
- Publication number
- CN114426584A CN114426584A CN202210130380.5A CN202210130380A CN114426584A CN 114426584 A CN114426584 A CN 114426584A CN 202210130380 A CN202210130380 A CN 202210130380A CN 114426584 A CN114426584 A CN 114426584A
- Authority
- CN
- China
- Prior art keywords
- fusion protein
- fragment
- amino acid
- tshr
- specific
- 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
- 210000003719 b-lymphocyte Anatomy 0.000 title claims abstract description 54
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 35
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 35
- 208000023275 Autoimmune disease Diseases 0.000 claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 4
- 239000012634 fragment Substances 0.000 claims description 24
- 210000004027 cell Anatomy 0.000 claims description 18
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 15
- 229920001184 polypeptide Polymers 0.000 claims description 13
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- 230000004048 modification Effects 0.000 claims description 3
- 238000012986 modification Methods 0.000 claims description 3
- 108020004707 nucleic acids Proteins 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 claims description 2
- 150000007523 nucleic acids Chemical class 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 7
- 102100026559 Filamin-B Human genes 0.000 claims 4
- 101000913551 Homo sapiens Filamin-B Proteins 0.000 claims 4
- 108010036012 Iodide peroxidase Proteins 0.000 claims 4
- 102000009843 Thyroglobulin Human genes 0.000 claims 4
- 108010034949 Thyroglobulin Proteins 0.000 claims 4
- 102000014267 Thyroid peroxidases Human genes 0.000 claims 4
- 229960002175 thyroglobulin Drugs 0.000 claims 4
- 238000009007 Diagnostic Kit Methods 0.000 claims 1
- 102000015486 thyroid-stimulating hormone receptor activity proteins Human genes 0.000 claims 1
- 108040006218 thyroid-stimulating hormone receptor activity proteins Proteins 0.000 claims 1
- 239000000427 antigen Substances 0.000 abstract description 10
- 102000036639 antigens Human genes 0.000 abstract description 10
- 108091007433 antigens Proteins 0.000 abstract description 10
- 238000003745 diagnosis Methods 0.000 abstract 1
- 210000002865 immune cell Anatomy 0.000 abstract 1
- 210000004408 hybridoma Anatomy 0.000 description 13
- 101150098159 TSHR gene Proteins 0.000 description 12
- 150000001413 amino acids Chemical group 0.000 description 11
- 230000003248 secreting effect Effects 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 6
- 210000004180 plasmocyte Anatomy 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 4
- 238000001976 enzyme digestion Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 210000001806 memory b lymphocyte Anatomy 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- HBZBAMXERPYTFS-SECBINFHSA-N (4S)-2-(6,7-dihydro-5H-pyrrolo[3,2-f][1,3]benzothiazol-2-yl)-4,5-dihydro-1,3-thiazole-4-carboxylic acid Chemical compound OC(=O)[C@H]1CSC(=N1)c1nc2cc3CCNc3cc2s1 HBZBAMXERPYTFS-SECBINFHSA-N 0.000 description 3
- 102000007079 Peptide Fragments Human genes 0.000 description 3
- 108010033276 Peptide Fragments Proteins 0.000 description 3
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 210000000649 b-lymphocyte subset Anatomy 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 210000001948 pro-b lymphocyte Anatomy 0.000 description 3
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 239000004695 Polyether sulfone Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 208000010928 autoimmune thyroid disease Diseases 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000008030 elimination Effects 0.000 description 2
- 238000003379 elimination reaction Methods 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 229920006393 polyether sulfone Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 102100031585 ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010002556 Ankylosing Spondylitis Diseases 0.000 description 1
- 230000024704 B cell apoptotic process Effects 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 208000008439 Biliary Liver Cirrhosis Diseases 0.000 description 1
- 208000033222 Biliary cirrhosis primary Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- GUTLYIVDDKVIGB-OUBTZVSYSA-N Cobalt-60 Chemical compound [60Co] GUTLYIVDDKVIGB-OUBTZVSYSA-N 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108091006020 Fc-tagged proteins Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 208000024869 Goodpasture syndrome Diseases 0.000 description 1
- 206010072579 Granulomatosis with polyangiitis Diseases 0.000 description 1
- 208000001204 Hashimoto Disease Diseases 0.000 description 1
- 208000030836 Hashimoto thyroiditis Diseases 0.000 description 1
- 101000777636 Homo sapiens ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010029240 Neuritis Diseases 0.000 description 1
- 206010034277 Pemphigoid Diseases 0.000 description 1
- 201000011152 Pemphigus Diseases 0.000 description 1
- 208000031845 Pernicious anaemia Diseases 0.000 description 1
- 206010036105 Polyneuropathy Diseases 0.000 description 1
- 208000012654 Primary biliary cholangitis Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 208000021386 Sjogren Syndrome Diseases 0.000 description 1
- 208000018359 Systemic autoimmune disease Diseases 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 229940125644 antibody drug Drugs 0.000 description 1
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000016644 chronic atrophic gastritis Diseases 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000004540 complement-dependent cytotoxicity Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 230000003090 exacerbative effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 210000003297 immature b lymphocyte Anatomy 0.000 description 1
- 230000008004 immune attack Effects 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 206010028417 myasthenia gravis Diseases 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000005868 ontogenesis Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 201000001976 pemphigus vulgaris Diseases 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 210000003720 plasmablast Anatomy 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 201000006292 polyarteritis nodosa Diseases 0.000 description 1
- 208000019629 polyneuritis Diseases 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000003146 transient transfection Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 208000035408 type 1 diabetes mellitus 1 Diseases 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
- C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/14—Drugs for disorders of the endocrine system of the thyroid hormones, e.g. T3, T4
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01008—Iodide peroxidase (1.11.1.8)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
Abstract
本发明公开了一种能够靶向清除特异性B淋巴细胞的融合蛋白及其应用,涉及生物技术领域。该融合蛋白包括B淋巴细胞结合结构域,以及与B淋巴细胞结合结构域连接的抗体Fc端,B淋巴细胞结合结构域具有与特异性B淋巴细胞结合的特性。该融合蛋白可以与被自身抗原激活的特异性B淋巴细胞结合,通过Fc端激活免疫细胞及分子,从而靶向清除特异性B淋巴细胞;该融合蛋白可应用于制备相应的药物及试剂盒,用于自身免疫疾病诊疗领域。
Description
技术领域
本发明涉及生物技术领域,具体而言,涉及一种融合蛋白及其应用。
背景技术
自身免疫性疾病是一种由自身抗原激活B淋巴细胞,产生自身抗体而导致对自身组织免疫攻击而产生的疾病,B淋巴细胞在自身免疫性疾病的发生发展过程中发挥重要作用。自身免疫性疾病的主要发病基础之一是产生自身抗体的自身反应性B淋巴细胞,因而B淋巴细胞清除疗法是一种重要的治疗方法。
人类B细胞主要在骨髓中发育,成熟后进入外周血,在其个体发育过程中,依次明确的表面抗原和DNA重排情况可分为:祖B细胞、前B细胞、未成熟B细胞、过渡B细胞、初始B细胞、记忆细胞、原浆细胞和浆细胞等。靶标一般为B细胞表面抗原,包括CD19(B细胞全生命周期表达)、CD20(Pro-B和浆细胞不表达)、CD38(浆母细胞,浆细胞表达)和CD138(浆细胞表达)等。目前B淋巴细胞清除策略是利用抗体药物靶向结合B细胞亚群的特定抗原,通过抗体依赖细胞介导的细胞毒性作用或补体依赖的细胞毒性作用介导B淋巴细胞裂解,从而清除B淋巴细胞,或者通过靶向B淋巴细胞存活需要的细胞因子,诱导B细胞凋亡,达到清除B淋巴细胞亚群的目的。
然而,目前上述方法清除的是特定B淋巴细胞亚群,这些B淋巴细胞亚群在调节免疫,在抗感染及抗肿瘤等方面发挥重要作用。因此,上述清除特定B淋巴细胞亚群的治疗药物存在着加剧免疫紊乱、导致感染及肿瘤发生风险等副作用。可见,如果能够特异性清除被自身抗原激活而产生自身特异性抗体的B淋巴细胞,而对其他B淋巴细胞亚群不产生影响,将极大提高自身免疫病的疗效和安全性。
鉴于此,特提出本发明。
发明内容
本发明的目的在于提供一种人工改造的该融合蛋白,可以与被特异性抗原激活的特异性B淋巴细胞结合,对特异性B淋巴细胞进行靶向清除。
本发明的另一目的在于提供一种细胞。
本发明的另一目的在于提供一种组合物。
本发明的另一目的在于提供上述细胞的应用。
本发明是这样实现的:
一种融合蛋白,其特征在于,其包括B淋巴细胞结合结构域、以及与B淋巴细胞结合结构域连接的抗体Fc端,B细胞结合结构域具有与被特异性抗原激活的特异性B淋巴细胞结合的特性。
进一步地,在本发明的一些实施方案中,所述特异性B淋巴细胞,是指能够被任意一种特异性抗原激活产生相应的特异性抗体的B淋巴细胞。
但需要说明的是,本发明的B细胞包括浆细胞和记忆B细胞。针对所述特异性B淋巴细胞应用到本发明中均属于本发明的保护范围。
当然,需要说明的是,特异性抗原,可以是任意能够诱导产生特异性抗体的所有动物来源的抗原。
进一步地,在本发明的一些实施方案中,TSHr片段包含SEQ ID NO.1序列中如表1所列的一个及多个氨基酸序列;TPO多肽片段包含SEQ ID NO.2序列中如表2所列的一个及多个氨基酸序列;TG多肽片段包含SEQ ID NO.3序列中如表3所列的一个及多个氨基酸序列。
含有表1的肽段可以靶向分泌TSHr抗体的特异性B淋巴细胞及记忆B淋巴细胞;含有表2的肽段可以靶向分泌TPO抗体的特异性B淋巴细胞及记忆B淋巴细胞;含有表3的肽段可以靶向分泌TG抗体的特异性B淋巴细胞及记忆B淋巴细胞。用于靶向清除产生甲状腺自身抗体的特异性B细胞,治疗自身免疫性甲状腺疾病。
该融合蛋白包含关键氨基酸序列,可结合于产生相应抗体的特异性B淋巴细胞,靶向清除特异性B淋巴细胞,用于治疗自身免疫性甲状腺疾病。
另一方面,本发明提供了一种细胞,其含有分离的核酸且表达有如上所述的融合蛋白。
另一方面,本发明提供了一种组合物,其包括如上所述的细胞以及药学上可接受的辅料。
另一方面,本发明提供了如上所述的细胞在制备治疗自身免疫疾病的药物中的应用。
例如,自身免疫疾病可以是慢性淋巴性甲状腺炎、甲状腺功能亢进、胰岛素依赖型糖尿病、重症肌无力、慢性溃疡性结肠炎、恶性贫血伴慢性萎缩性胃炎、肺出血肾炎综合征、寻常天疱疮、类天疱疮、原发性胆汁性肝硬变、多发性脑脊髓硬化症、急性特发性多神经炎等;还有系统性自身免疫病如系统性红斑狼疮、口眼干燥综合征、类风湿性关节炎、强直性脊柱炎、硬皮病、结节性多动脉炎或Wegener肉芽肿病等。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,应当理解,以下附图仅示出了本发明的某些实施例,因此不应被看作是对范围的限定,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他相关的附图。
图1为TSHr胞外段Fc融合蛋白特异性清除特异性B淋巴细胞示意图。
图2是:流式细胞仪分析荧光素偶联融合蛋白与杂交瘤细胞的结合。结果: TSHr-Fc融合蛋白与对照组杂交瘤细胞结合率15%,而与分泌抗TSHr抗体的杂 交瘤细胞结合率达93.3%,上述结果说明,TSHr-Fc融合蛋白可以与分泌抗TSHr 抗体的杂交瘤细胞特异性结合。
图3是:尾静脉输注杂交瘤细胞,该细胞可分泌抗TSHr抗体及表达荧光素酶, 5天后腹腔注射荧光素酶底物,观察杂交瘤细胞在小鼠体内的生长。融合蛋白组, 注射20ugTSHr-Fc融合蛋白,对照组注射相同剂量白蛋白。每隔5天采集活体 荧光成像数据,荧光信号与杂交瘤数量成正比。结果表明,对照小鼠体内肿瘤细 胞增殖迅速,而融合蛋白的小鼠体内杂交瘤细胞至第14天被完全清除,表明 TSHr-Fc融合蛋白可有效清除分泌抗TSHr抗体特异性B细胞。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
实施例1
本实施例以靶向特异性产生人TSHr自身抗体的B细胞为例,对本实施例的融合蛋白的结构进行说明,如下:
该融合蛋白包括特异性B淋巴细胞结合结构域、以及与所述结合结构域连接的Fc端,所述B细胞结合结构域是一段含有TSHr胞外段(21-413aa)多肽片段,具有与分泌抗TSHr抗体的特异性B淋巴细胞结合的特性。
获取TSHr的氨基酸序列信息及蛋白结构信息,选取第一个胞外区氨基酸对应的核苷酸序列,在其3’端加上Fc标签后,进行全基因合成后,制备模板片段,采用BamHI-XhoI进行双酶切后,回收目标基因DNA片段。将真核表达载体pcDNA3.1+使用BamHI-XhoI进行双酶切,进行琼脂糖凝胶电泳后,回收酶切后的载体,与上述酶切后的片段进行T4 DNA连接酶连接,转化至大肠杆菌感受态TOP10细胞中,提取质粒DNA后,进行Sanger测序验证,293F细胞的瞬时转染,收集培养基上清,用0.45um的PES材质滤膜将过滤后,转至无菌离心管中,使用蠕动泵将上清泵入Protein A柱,使用pH 3.0的甘氨酸溶液将TSHR-Fc蛋白从Protein A柱中洗脱下来,并迅速采用pH值9.0的Tris缓冲液中和至pH=7.0,采用截留分子量为30kDa的超滤管进行超滤,将缓冲液置换为PBS缓冲液,蛋白分装后置于-80°C保存。
实施例2 含有关键氨基酸序列的TSHr、TPO、TG多肽片段的获取
以抗人TSHr多克隆抗体作为靶标,对噬菌体展示随机十二肽库进行筛选。通过ELISA鉴定筛选克隆的结合特性,并对阳性克隆提取DNA进行测序分析。结果显示,经3轮生物淘洗后,目标噬菌体得到350倍富集.随机挑选80个克隆进行ELISA鉴定,其中有75个噬菌体克隆可以与抗人TSHr多克隆抗体特异性结合。测序分析发现,这75个克隆带有16种氨基酸序列(表1),与TSHr蛋白序列同源,为含有关键氨基酸序列的TSHr多肽片段,可以与分泌抗TSHr抗体的特异性B淋巴细胞结合。同样地,我们鉴定了29种含有关键氨基酸序列的TPO多肽片段(表2),以及84种含有关键氨基酸序列的TG多肽片段(表3)。
实验例:
1.TSHr-Fc融合蛋白对靶细胞体内杀伤实验
(1)6周龄雌性NSG小鼠10只,平均体重18.3-23.5g,饲养于恒温恒湿的独立通风盒内,饲养室温度22-23°C,湿度56-60%,10-20次/小时换气,昼夜明暗交替时间12h/12h;持续供给经钴60放射灭菌鼠全价颗粒饲料,不限量自由摄取,饮用自来水(高压蒸汽灭菌后使用),饮水瓶不间断供水,自由摄取。
(2)Nalm6-Luc细胞培养在含10% 胎牛血清的DMEM培养液中。该细胞由我们实验室自行制备保种,收集指数生长期的细胞,PBS重悬至适合浓度用于接种。
(3)所有小鼠经尾静脉输注Nalm6-Luc细胞,每只接种5*106个产生TSHr抗体的杂交瘤细胞,小鼠连续饲养5天后,腹腔注射荧光素酶底物D-luciferin,进行预实验观察肿瘤在小鼠体内的生长情况。根据肿瘤大小平均随机分为2组。
(4)其中一组小鼠为处理组,每只小鼠注射20ug TSHr-Fc融合蛋白;另一组小鼠为对照组。每隔5天对小鼠腹腔注射荧光素酶底物D-luciferin同时使用气体麻醉机对小鼠进行麻醉,采集肿瘤活体成像数据。
实验结果
2.1 TSHr-Fc融合蛋白与杂交瘤细胞的结合分析
取5*106个分泌抗TSHr抗体的杂交瘤细胞及相同数量的对照细胞,分别与异硫氰酸荧光素(FITC)偶联的TSHr-Fc融合蛋白室温孵育30分钟,用生理盐水洗涤三次后,使用流式细胞仪分析杂交瘤细胞膜抗TSHr抗体与TSHr-Fc融合蛋白的结合情况。如图2所示,分泌抗TSHr抗体的杂交瘤细胞可以与TSHr-Fc融合蛋白结合。
小鼠体内TSHr-Fc融合蛋白靶向杀伤特异性B淋巴细胞作用
在小鼠体内移植分泌抗TSHr的杂交瘤细胞,然后输注TSHr-Fc融合蛋白,定期观察小鼠体内肿瘤细胞的生长情况,结果表明,对照小鼠体内肿瘤细胞增殖迅速,而输注TSHr-Fc融合蛋白的小鼠体内肿瘤细胞随着时间的推移,完全被清除,表明融合蛋白在体内亦可有效的靶向清除小鼠体内分泌抗TSHr抗体特异性B淋巴细胞(图3)。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
SEQIDNO.1
MRPADLLQLVLLLDLPRDLGGMGCSSPPCECHQEEDFRVTCKDIQRIPSLPPSTQTLKLIETHLRTIPSHAFSNLPNISRIYVSIDVTLQQLESHSFYNLSKVTHIEIRNTRNLTYIDPDALKELPLLKFLGIFNTGLKMFPDLTKVYSTDIFFILEITDNPYMTSIPVNAFQGLCNETLTLKLYNNGFTSVQGYAFNGTKLDAVYLNKNKYLTVIDKDAFGGVYSGPSLLDVSQTSVTALPSKGLEHLKELIARNTWTLKKLPLSLSFLHLTRADLSYPSHCCAFKNQKKIRGILESLMCNESSMQSLRQRKSVNALNSPLHQEYEENLGDSIVGYKEKSKFQDTHNNAHYYVFFEEQEDEIIGFGQELKNPQEETLQAFDSHYDYTICGDSEDMVCTPKSDEFNPCEDIMGYKFLRIVVWFVSLLALLGNVFVLLILLTSHYKLNVPRFLMCNLAFADFCMGMYLLLIASVDLYTHSEYYNHAIDWQTGPGCNTAGFFTVFASELSVYTLTVITLERWYAITFAMRLDRKIRLRHACAIMVGGWVCCFLLALLPLVGISSYAKVSICLPMDTETPLALAYIVFVLTLNIVAFVIVCCCYVKIYITVRNPQYNPGDKDTKIAKRMAVLIFTDFICMAPISFYALSAILNKPLITVSNSKILLVLFYPLNSCANPFLYAIFTKAFQRDVFILLSKFGICKRQAQAYRGQRVPPKNSTDIQVQKVTHEMRQGLHNMEDVYELIENSHLTPKKQGQISEEYMQTVL
SEQIDNO.2 MRALAVLSVTLVMACTEAFFPFISRGKELLWGKPEESRVSSVLEESKRLVDTAMYATMQRNLKKRGILSPAQLLSFSKLPEPTSGVIARAAEIMETSIQAMKRKVNLKTQQSQHPTDALSEDLLSIIANMSGCLPYMLPPKCPNTCLANKYRPITGACNNRDHPRWGASNTALARWLPPVYEDGFSQPRGWNPGFLYNGFPLPPVREVTRHVIQVSNEVVTDDDRYSDLLMAWGQYIDHDIAFTPQSTSKAAFGGGADCQMTCENQNPCFPIQLPEEARPAAGTACLPFYRSSAACGTGDQGALFGNLSTANPRQQMNGLTSFLDASTVYGSSPALERQLRNWTSAEGLLRVHARLRDSGRAYLPFVPPRAPAACAPEPGIPGETRGPCFLAGDGRASEVPSLTALHTLWLREHNRLAAALKALNAHWSADAVYQEARKVVGALHQIITLRDYIPRILGPEAFQQYVGPYEGYDSTANPTVSNVFSTAAF RFGHATIHPLVRRLDASFQEHPDLPGLWLHQAFFSPWTLLRGGGLDPLIRGLLARPAKLQVQDQLMNEELTERLFVLSNSSTLDLASINLQRGRDHGLPGYNEWREFCGLPRLETPADLSTAIASRSVADKILDLYKHPDNIDVWLGGLAENFLPRARTGPLFACLIGKQMKALRDGDWFWWENSHVFTDAQRRELEKHSLSRVICDNTGLTRVPMDAFQVGKFPEDFESCDSITGMNLEAWRETFPQDDKCGFPESVENGDFVHCEESGRRVLVYSCRHGYELQGREQLTCTQEGWDFQPPLCKDVNECADGAHPPCHASARCRNTKGGFQCLCADPYELGDDGRTCVDSGRLPRVTWISMSLAALLIGGFAGLTSTVICRWTRTGTKSTLPISETGGGTPELRCGKHQAVGTSPQRAAAQDSEQESAGMEGRDTHRLPRAL
SEQIDNO.3 MALVLEIFTLLASICWVSANIFEYQVDAQPLRPCELQRETAFLKQADYVPQCAEDGSFQTVQCQNDGRSCWCVGANGSEVLGSRQPGRPVACLSFCQLQKQQILLSGYINSTDTSYLPQCQDSGDYAPVQCDVQQVQCWCVDAEGMEVYGTRQLGRPKRCPRSCEIRNRRLLHGVGDKSPPQCSAEGEFMPVQCKFVNTTDMMIFDLVHSYNRFPDAFVTFSSFQRRFPEVSGYCHCADSQGRELAETGLELLLDEIYDTIFAGLDLPSTFTETTLYRILQRRFLAVQSVISGRFRCPTKCEVERFTATSFGHPYVPSCRRNGDYQAVQCQTEGPCWCVDAQGKEMHGTRQQGEPPSCAEGQSCASERQQALSRLYFGTSGYFSQHDLFSSPEKRWASPRVARFATSCPPTIKELFVDSGLLRPMVEGQSQQFSVSENLLKEAIRAIFPSRGLARLALQFTTNPKRLQQNLFGGKFLVNVGQFNLSGALG TRGTFNFSQFFQQLGLASFLNGGRQEDLAKPLSVGLDSNSSTGTPEAAKKDGTMNKPTVGSFGFEINLQENQNALKFLASLLELPEFLLFLQHAISVPEDVARDLGDVMETVLSSQTCEQTPERLFVPSCTTEGSYEDVQCFSGECWCVNSWGKELPGSRVRGGQPRCPTDCEKQRARMQSLMGSQPAGSTLFVPACTSEGHFLPVQCFNSECYCVDAEGQAIPGTRSAIGKPKKCPTPCQLQSEQAFLRTVQALLSNSSMLPTLSDTYIPQCSTDGQWRQVQCNGPPEQVFELYQRWEAQNKGQDLTPAKLLVKIMSYREAASGNFSLFIQSLYEAGQQDVFPVLSQYPSLQDVPLAALEGKRPQPRENILLEPYLFWQILNGQLSQYPGSYSDFSTPLAHFDLRNCWCVDEAGQELEGMRSEPSKLPTCPGSCEEAKLRVLQFIRETEEIVSASNSSRFPLGESFLVAKGIRLRNEDLGLPPLFPPREAFAEQFLRGSDYAIRLAAQSTLSFYQRRRFSPDDSAGASALLRSGPYMPQCDAFGSWEPVQCHAGTGHCWCVDEKGGFIPGSLTARSLQIPQCPTTCEKSRTSGLLSSWKQARSQENPSPKDLFVPACLETGEYARLQASGAGTWCVDPASGEELRPGSSSSAQCPSLCNVLKSGVLSRRVSPGYVPACRAEDGGFSPVQCDQAQGSCWCVMDSGEEVPGTRVTGGQPACESPRCPLPFNASEVVGGTILCETISGPTGSAMQQCQLLCRQGSWSVFPPGPLICSLESGRWESQLPQPRACQRPQLWQTIQTQGHFQLQLPPGKMCSADYADLLQTFQVFILDELTARGFCQIQVKTFGTLVSIPVCNNSSVQVGCLTRERLGVNVTWKSRLEDIPVASLPDLHDIERALVGKDLLGRFTDLIQSGSFQLHLDSKTFPAETIRFLQGDHFGTSPRTWFGCSEGFYQVLTSEASQDGLGCVKCPEGSYSQDEECIPCPVGFYQEQAGSLACVPCPVGRTTISAGAFSQTHCVTDCQRNEAGLQCDQNGQYRASQKDRGSGKAFCVDGEGRRLPWWETEAPLEDSQCLMMQKFEKVPESKVIFDANAPVAVRSKVPDSEFPVMQCLTDCTEDEACSFFTVSTTEPEISCDFYAWTSDNVACMTSDQKRDALGNSKATSFGSLRCQVKVRSHGQDSPAVYLKKGQGSTTTLQKRFEPTGFQNMLSGLYNPIVFSASGANLTDAHLFCLLACDRDLCCDGFVLTQVQGGAIICGLLSSPSVLLCNVKDWMDPSEAWANATCPGVTYDQESHQVILRLGDQEFIKSLTPLEGTQDTFTNFQQVYLWKDSDMGSRPESMGCRKDTVPRPASPTEAGLTTELFSPVDLNQVIVNGNQSLSSQKHWLFKHLFSAQQANLWCLSRCVQEHSFCQLAEITESASLYFTCTLYPEAQVCDDIMESNAQGCRLILPQMPKALFRKKVILEDK
VKNFYTRLPFQKLMGISIRNKVPMSEKSISNGFFECERRCDADPCCTGFGFLNVSQLKGGEVTCLTLNSLGIQMCSEENGGAWRILDCGSPDIEVHTYPFGWYQKPIAQNNAPSFCPLVVLPSLTEKVSLDSWQSLALSSVVVDPSIRHFDVAHVSTAATSNFSAVRDLCLSECSQHEACLITTLQTQPGAVRCMFYADTQSCTHSLQGQNCRLLLREEATHIYRKPGISLLSYEASVPSVPISTHGRLLGRSQAIQVGTSWKQVDQFLGVPYAAPPLAERRFQAPEPLNWTGSWDASKPRASCWQPGTRTSTSPGVSEDCLYLNVFIPQNVAPNASVLVFFHNTMDREESEGWPAIDGSFLAAVGNLIVVTASYRVGVFGFLSSGSGEVSGNWGLLDQVAALTWVQTHIRGFGGDPRRVSLAADRGGADVASIHLLTARATNSQLFRRAVLMGGSALSPAAVISHERAQQQAIALAKEVSCPMSSSQEVVSCLRQKPANVLNDAQTKLLAVSGPFHYWGPVIDGHFLREPPARALKRSLWVEVDLLIGSSQDDGLINRAKAVKQFEESRGRTSSKTAFYQALQNSLGGEDSDARVEAAATWYYSLEHSTDDYASFSRALENATRDYFIICPIIDMASAWAKRARGNVFMYHAPENYGHGSLELLADVQFALGLPFYPAYEGQFSLEEKSLSLKIMQYFSHFIRSGNPNYPYEFSRKVPTFATPWPDFVPRAGGENYKEFSELLPNRQGLKKADCSFWSKYISSLKTSADGAKGGQSAESEEEELTAGSGLREDLLSLQEPGSKTYSK
Claims (10)
1.一种融合蛋白,其特征在于,其包括B淋巴细胞结合结构域,以及与B淋巴细胞结合结构域连接的抗体Fc端,B淋巴细胞结合结构域具有与特异性B淋巴细胞结合的特性。
2.根据权利要求1所述的融合蛋白,其特征在于,所述的B淋巴细胞结合结构域为自身抗原片段。
3.根据权利要求1所述的融合蛋白,其特征在于,所述抗体Fc端为人源IgG1、IgG2、IgG3、IgG4的FC端及其修饰体及突变体。
4.根据权利要求2所述的自身抗原片段,是指甲状腺自身抗原片段,包括:促甲状腺素受体(TSHr)、甲状腺过氧化酶(TPO)及甲状腺球蛋白(TG)片段。
5.根据权利要求4所述,其特征在于,所述甲状腺自身抗原片段是指含有B淋巴细胞结合结构域关键氨基酸序列的多肽片段。
6.根据权利要求5所述,含有B淋巴细胞结合结构域关键氨基酸序列的多肽片段,是指能够与TSHr抗体、TPO抗体及TG抗体结合的含有关键氨基酸序列的甲状腺自身抗原片段。
7.根据权利要求6所述的含有关键氨基酸序列的甲状腺自身抗原片段,其特征在于,TSHr片段包含SEQ ID NO.1序列中如表1所列的一个及多个氨基酸序列;TPO多肽片段包含SEQ ID NO.2序列中如表2所列的一个及多个氨基酸序列;TG多肽片段包含SEQ ID NO.3序列中如表3所列的一个及多个氨基酸序列。
8.一种细胞,其特征在于,其含有编码表达权利要求1-7所述的融合蛋白的分离的核酸及载体。
9.一种组合物,其特征在于,其包括权利要求8所述的细胞以及药学上可接受的辅料。
10.权利要求1及权利要求8-9所述的融合蛋白及细胞在制备自身免疫疾病治疗药物及诊断试剂盒中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210130380.5A CN114426584A (zh) | 2022-02-11 | 2022-02-11 | 一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210130380.5A CN114426584A (zh) | 2022-02-11 | 2022-02-11 | 一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114426584A true CN114426584A (zh) | 2022-05-03 |
Family
ID=81312616
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210130380.5A Pending CN114426584A (zh) | 2022-02-11 | 2022-02-11 | 一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114426584A (zh) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016166139A1 (en) * | 2015-04-14 | 2016-10-20 | Eberhard Karls Universität Tübingen | Bispecific fusion proteins for enhancing immune responses of lymphocytes against tumor cells |
CN107098978A (zh) * | 2017-05-05 | 2017-08-29 | 中国药科大学 | 一种抗肿瘤和免疫增强双重功效融合蛋白 |
CN111372600A (zh) * | 2017-03-24 | 2020-07-03 | 奥菲斯生物科学公司 | 治疗自身免疫性疾病的PantId |
-
2022
- 2022-02-11 CN CN202210130380.5A patent/CN114426584A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016166139A1 (en) * | 2015-04-14 | 2016-10-20 | Eberhard Karls Universität Tübingen | Bispecific fusion proteins for enhancing immune responses of lymphocytes against tumor cells |
CN111372600A (zh) * | 2017-03-24 | 2020-07-03 | 奥菲斯生物科学公司 | 治疗自身免疫性疾病的PantId |
CN107098978A (zh) * | 2017-05-05 | 2017-08-29 | 中国药科大学 | 一种抗肿瘤和免疫增强双重功效融合蛋白 |
Non-Patent Citations (6)
Title |
---|
MARCINKOWSKI P等: "thyrotropin receptor isoform 1 precursor [Homo sapiens]NP_000360.2", GENBANK, pages 3 - 4 * |
MATANA A等: "thyroglobulin precursor [Homo sapiens].NP_003226.4", GENBANK, pages 6 - 7 * |
RAMOS-LEVÍ AM等: "Pathogenesis of thyroid autoimmune disease: the role of cellular mechanisms", 《ENDOCRINOLOGIA Y NUTRICION》, vol. 63, no. 8, pages 424 * |
WILLIAMS DE等: "thyroid peroxidase isoform a precursor [Homo sapiens].NP_000538.3", GENBANK, pages 3 * |
高洁等: "AChR-IgG Fc 段融合蛋白真核表达载体的构建及表达", 《中国神经免疫学和神经病 学杂志》, vol. 17, no. 3, pages 188 * |
高洁等: "AChR-IgG Fc段融合蛋白真核表达载体的构建及表达", 《中国神经免疫学和神经病学杂志》, vol. 17, no. 3, pages 188 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107217042A (zh) | 一种生产无岩藻糖基化蛋白的基因工程细胞系及其建立方法 | |
US20180369285A1 (en) | Specific Chimeric Antigen Receptor T Cells Targeting To NKG2DL, Its Preparation Method and Application Thereof | |
CN110835371A (zh) | 抗ccr8单克隆抗体及其应用 | |
JP6757712B2 (ja) | 幹細胞増強治療法 | |
CN102482324A (zh) | 能够抑制自身免疫、炎症和癌症进程的新型趋化因子结合多肽 | |
US8404803B2 (en) | Cancer-associated antigen analogue peptides and uses thereof | |
JP4857396B1 (ja) | 融合蛋白質 | |
CN111909271B (zh) | 一种基于单域抗体的bcma嵌合抗原受体及其应用 | |
CN103826659A (zh) | 用于治疗的靶向神经肌肉接头 | |
CN109652378A (zh) | 一种功能增强的通用型car-t细胞及其制备方法和用途 | |
CN108463243A (zh) | 重组人c1酯酶抑制剂及其用途 | |
CN110312527A (zh) | 用于干细胞移植的非基因毒性预处理方案 | |
CN114853905B (zh) | 一种基因修饰nk细胞与抗体联合使用治疗肿瘤方案 | |
TWI758884B (zh) | 含人白細胞介素10和Fc片段的融合蛋白及其醫藥用途 | |
CN102470169A (zh) | 自身抗体产生抑制剂 | |
CN111542547B (zh) | 对bdca2抗原具有特异性的嵌合抗原受体 | |
CN114426584A (zh) | 一种靶向清除特异性b淋巴细胞的融合蛋白及其应用 | |
JPH05501118A (ja) | 合成ポリ―Igレセプター,レセプター―抗体複合体、その生産および用途 | |
CN114702595A (zh) | 一种靶向杀伤特异性b淋巴细胞的融合蛋白及其应用 | |
CA3115139A1 (en) | Compositions and methods regarding engineered and non-engineered .gamma..delta.-t cells for treatment of hematological tumors | |
CN102559636A (zh) | 用于白血病和自身免疫疾病的抗体融合蛋白及其制备方法 | |
CN110177802A (zh) | 多聚化stradomer GL-2045的制造优化 | |
JP7439280B2 (ja) | キメラ抗原受容体の最適化 | |
CN114621355A (zh) | 一种靶向特异性b淋巴细胞的嵌合抗原受体及其应用 | |
CN103524625B (zh) | 一种新型诱导抗体产生的方法 |
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 | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20220503 |