CN107881142A - 一种杂交瘤细胞无血清培养基 - Google Patents

一种杂交瘤细胞无血清培养基 Download PDF

Info

Publication number
CN107881142A
CN107881142A CN201711319522.8A CN201711319522A CN107881142A CN 107881142 A CN107881142 A CN 107881142A CN 201711319522 A CN201711319522 A CN 201711319522A CN 107881142 A CN107881142 A CN 107881142A
Authority
CN
China
Prior art keywords
acid
sodium
chloride
vitamin
culture medium
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.)
Withdrawn
Application number
CN201711319522.8A
Other languages
English (en)
Inventor
刘才明
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chengdu Source Biotechnology Co Ltd
Original Assignee
Chengdu Source Biotechnology Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Chengdu Source Biotechnology Co Ltd filed Critical Chengdu Source Biotechnology Co Ltd
Priority to CN201711319522.8A priority Critical patent/CN107881142A/zh
Publication of CN107881142A publication Critical patent/CN107881142A/zh
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • C12N5/0018—Culture media for cell or tissue culture
    • C12N5/0037—Serum-free medium, which may still contain naturally-sourced components
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • C12N2500/14—Calcium; Ca chelators; Calcitonin
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/12—Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
    • C12N2500/16—Magnesium; Mg chelators
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/32—Amino acids
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/34—Sugars
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/36—Lipids
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/38—Vitamins
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/40—Nucleotides, nucleosides, bases
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/44—Thiols, e.g. mercaptoethanol
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C12N2500/46—Amines, e.g. putrescine
    • 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
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/33—Insulin
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/39—Steroid hormones
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/39—Steroid hormones
    • C12N2501/392—Sexual steroids
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/998—Proteins not provided for elsewhere
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/999—Small molecules not provided for elsewhere

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Cosmetics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

本发明公开了一种杂交瘤细胞无血清培养基，包括如下原料：丙氨酸、精氨酸、天冬氨酸、天冬酰氨、胱氨酸、半胱氨酸、谷氨酰胺、谷氨酸、甘氨酸、组氨酸等，氯化钙、硫酸镁、氯化钾、磷酸二氢钾、磷酸氢二钠、碳酸氢钠、氢氧化钠、七水合硫酸亚铁、六水合氯化镁、硫酸铜、丙酮酸钠、二氯化锰、亚硒酸钠、钼酸铵等，尼克酰胺、对氨基苯甲酸、叶酸、核糖、尿嘧啶、硫辛酸、亚油酸L、胆固醇、地塞米松、雌二醇孕酮、睾酮等。本发明培养基，完全不含有血清成份；生产的抗体纯度大幅度提高；该培养基同时具有高扩增速度及高抗体表达能力；该培养基能长时间保持细胞活性，杂交瘤细胞培养时间大大延长，可以大大延长生产时间，提高抗体产量。

Description

一种杂交瘤细胞无血清培养基
技术领域
本发明涉及一种细胞培养基，具体涉及一种杂交瘤细胞无血清培养基。
背景技术
细胞培养经历了几十年的发展，现广泛应用于生物、医疗技术、药品等领域，形成了庞大的产业。产品越来越细分化，以适应不同的目的需求。通过动物工程细胞培养获得蛋白药物和抗体已经成为现代医药发展的重要方向。杂交瘤细胞培养获取抗体作为药物生产非常重要的手段，已经广泛用于单抗、多抗药物的生产。但杂交瘤的培养生产工艺相对比较复杂。生产工艺中最重要一环杂交瘤培养基开发难度极大，目前国内多数生产厂家采用的工艺都有动物血清添加，而这些成份的添加往往可能带来外源IgG导致目标抗体在分离纯化上难度的增加，对产品的品质造成影响。
发明内容
本发明所要解决的技术问题是杂交瘤的培养生产工艺相对比较复杂，生产工艺中最重要一环杂交瘤培养基开发难度极大，目前国内多数生产厂家采用的工艺都有动物血清添加，而这些成份的添加往往可能带来外源IgG导致目标抗体在分离纯化上难度的增加，对产品的品质造成影响，目的在于提供一种杂交瘤细胞无血清培养基，完全不含有血清成份；生产的抗体纯度大幅度提高；该培养基同时具有高扩增速度及高抗体表达能力；杂交瘤细胞不需要驯化，可以直接从含血清培养直接过渡到无血清培养基培养；该培养基能长时间保持细胞活性，杂交瘤细胞培养时间大大延长，可以大大延长生产时间，提高抗体产量。
本发明通过下述技术方案实现：
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸10-100mg/L、精氨酸10-100mg/L、天冬氨酸10-100mg/L、天冬酰氨10-100mg/L、胱氨酸10-100mg/L、半胱氨酸10-100mg/L、谷氨酰胺100-1000mg/L、谷氨酸10-100mg/L、甘氨酸10-100mg/L、组氨酸 10-100mg/L、羟脯氨酸10-100mg/L、异亮氨酸10-100mg/L、亮氨酸10-100mg/L、赖氨酸 10-100mg/L、蛋氨酸10-100mg/L、苯丙氨酸10-100mg/L、脯氨酸10-100mg/L、丝氨酸 10-100mg/L、苏氨酸10-100mg/L、色氨酸10-100mg/L、酪氨酸10-100mg/L、缬氨酸10-100mg/L、氯化钙10-100mg/L、硫酸镁10-100mg/L、氯化钾10-1000mg/L、磷酸二氢钾10-100mg/L、磷酸氢二钠10-100mg/L、碳酸氢钠0.1-5g/L、氢氧化钠0.1-5g/L、七水合硫酸亚铁10-100mg/L、六水合氯化镁10-100mg/L、硫酸铜10-100mg/L、丙酮酸钠10-100mg/L、二氯化锰10-100mg/L、亚硒酸钠10-100mg/L、钼酸铵0.001-100mg/L、丁酸钠10-100mg/L、丁二酸钠10-100mg/L、柠檬酸铁10-100mg/L、柠檬酸10-100mg/L、氯化钠1-10g/L、生物素1-100mg/L、氯化胆碱 1-100mg/L、酒石酸胆碱1-100mg/L、肌醇1-100mg/L、尼克酰胺1-100mg/L、D-泛酸半钙 1-100mg/L、吡哆醛1-100mg/L、硫胺素1-100mg/L、核黄素1-100mg/L、维生素B2 1-100mg/L、维生素D2 1-100mg/L、抗坏血酸1-100mg/L、维生素K2 1-100mg/L、维生素K3 1-100mg/L、维生素B-120.1-100mg/L、对氨基苯甲酸1-100mg/L、叶酸1-100mg/L、D-葡萄糖 1000-6000mg/L、胸苷10-100mg/L、谷胱苷肽还原型1-100mg/L、腐胺1-100mg/L、乙醇胺 1-100mg/L、β-巯基乙醇1-100mg/L、次黄嘌呤1-100mg/L、腺嘌呤1-100mg/L、盐酸鸟嘌呤 1-100mg/L、核糖1-100mg/L、尿嘧啶1-100mg/L、硫辛酸1-100mg/L、亚油酸1-100mL/L、胆固醇1-100mg/L、氨基多糖1-100mg/L、hepes 1-10g/L、胰岛素0.00001-10mg/L、氢化可的松 0.00001-10mg/L、地塞米松0.00001-10mg/L、雌二醇孕酮0.00001-10mg/L、睾酮 0.00001-10mg/L、酵母水解物10-1000mg/l、水解乳蛋白10-1000mg/l、酪蛋白水解物 10-1000mg/l。
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸30-100mg/L、精氨酸10-80mg/L、天冬氨酸10-70mg/L、天冬酰氨20-100mg/L、胱氨酸10-90mg/L、半胱氨酸50-100mg/L、谷氨酰胺300-1000mg/L、谷氨酸10-70mg/L、甘氨酸20-80mg/L、组氨酸 10-60mg/L、羟脯氨酸40-100mg/L、异亮氨酸20-100mg/L、亮氨酸10-60mg/L、赖氨酸 50-100mg/L、蛋氨酸30-90mg/L、苯丙氨酸40-100mg/L、脯氨酸20-60mg/L、丝氨酸20-60mg/L、苏氨酸20-60mg/L、色氨酸20-60mg/L、酪氨酸20-60mg/L、缬氨酸20-60mg/L、氯化钙 20-60mg/L、硫酸镁20-60mg/L、氯化钾300-1000mg/L、磷酸二氢钾30-70mg/L、磷酸氢二钠 30-70mg/L、碳酸氢钠0.1-2g/L、氢氧化钠2-5g/L、七水合硫酸亚铁30-70mg/L、六水合氯化镁30-70mg/L、硫酸铜10-90mg/L、丙酮酸钠30-70mg/L、二氯化锰40-100mg/L、亚硒酸钠 30-70mg/L、钼酸铵10-100mg/L、丁酸钠30-70mg/L、丁二酸钠30-70mg/L、柠檬酸铁10-50mg/L、柠檬酸30-70mg/L、氯化钠1-6g/L、生物素30-70mg/L、氯化胆碱20-70mg/L、酒石酸胆碱 30-60mg/L、肌醇40-80mg/L、尼克酰胺30-70mg/L、D-泛酸半钙30-50mg/L、吡哆醛10-70mg/L、硫胺素30-100mg/L、核黄素1-80mg/L、维生素B2 30-70mg/L、维生素D2 20-70mg/L、抗坏血酸30-80mg/L、维生素K2 10-70mg/L、维生素K3 10-60mg/L、维生素B-12 0.1-50mg/L、对氨基苯甲酸10-40mg/L、叶酸30-70mg/L、D-葡萄糖3000-6000mg/L、胸苷80-100mg/L、谷胱苷肽还原型10-50mg/L、腐胺30-70mg/L、乙醇胺30-70mg/L、β-巯基乙醇40-100mg/L、次黄嘌呤30-90mg/L、腺嘌呤30-70mg/L、盐酸鸟嘌呤30-70mg/L、核糖30-70mg/L、尿嘧啶 20-100mg/L、硫辛酸30-80mg/L、亚油酸15-60mL/L、胆固醇70-100mg/L、氨基多糖50-90mg/L、 hepes 3-8g/L、胰岛素0.00001-5mg/L、氢化可的松1-10mg/L、地塞米松0.00001-2mg/L、雌二醇孕酮0.00001-3mg/L、睾酮0.5-2mg/L、酵母水解物400-1000mg/l、水解乳蛋白 500-1000mg/l、酪蛋白水解物10-1000mg/l。
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸50mg/L、精氨酸40mg/L、天冬氨酸30mg/L、天冬酰氨50mg/L、胱氨酸40mg/L、半胱氨酸70mg/L、谷氨酰胺600mg/L、谷氨酸20mg/L、甘氨酸30mg/L、组氨酸50mg/L、羟脯氨酸70mg/L、异亮氨酸80mg/L、亮氨酸30mg/L、赖氨酸90mg/L、蛋氨酸60mg/L、苯丙氨酸50mg/L、脯氨酸40mg/L、丝氨酸50mg/L、苏氨酸40mg/L、色氨酸30mg/L、酪氨酸50mg/L、缬氨酸30mg/L、氯化钙40mg/L、硫酸镁35mg/L、氯化钾700mg/L、磷酸二氢钾60mg/L、磷酸氢二钠40mg/L、碳酸氢钠1g/L、氢氧化钠3g/L、七水合硫酸亚铁40mg/L、六水合氯化镁70mg/L、硫酸铜30mg/L、丙酮酸钠 40mg/L、二氯化锰50mg/L、亚硒酸钠40mg/L、钼酸铵5mg/L、丁酸钠70mg/L、丁二酸钠30mg/L、柠檬酸铁20mg/L、柠檬酸55mg/L、氯化钠3g/L、生物素40mg/L、氯化胆碱25mg/L、酒石酸胆碱60mg/L、肌醇40mg/L、尼克酰胺35mg/L、D-泛酸半钙40mg/L、吡哆醛10mg/L、硫胺素100mg/L、核黄素70mg/L、维生素B2 50mg/L、维生素D2 20mg/L、抗坏血酸60mg/L、维生素K2 50mg/L、维生素K3 20mg/L、维生素B-12 10mg/L、对氨基苯甲酸15mg/L、叶酸 60mg/L、D-葡萄糖4000mg/L、胸苷90mg/L、谷胱苷肽还原型30mg/L、腐胺50mg/L、乙醇胺45mg/L、β-巯基乙醇80mg/L、次黄嘌呤70mg/L、腺嘌呤45mg/L、盐酸鸟嘌呤50mg/L、核糖45mg/L、尿嘧啶55mg/L、硫辛酸60mg/L、亚油酸30mL/L、胆固醇80mg/L、氨基多糖 60mg/L、hepes 5g/L、胰岛素2mg/L、氢化可的松40mg/L、地塞米松0.5mg/L、雌二醇孕酮 1mg/L、睾酮1mg/L、酵母水解物600mg/l、水解乳蛋白800mg/l、酪蛋白水解物600mg/l。
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸10mg/L、精氨酸10mg/L、天冬氨酸10mg/L、天冬酰氨10mg/L、胱氨酸10mg/L、半胱氨酸10mg/L、谷氨酰胺100mg/L、谷氨酸10mg/L、甘氨酸10mg/L、组氨酸10mg/L、羟脯氨酸10mg/L、异亮氨酸10mg/L、亮氨酸10mg/L、赖氨酸10mg/L、蛋氨酸10mg/L、苯丙氨酸10mg/L、脯氨酸10mg/L、丝氨酸10mg/L、苏氨酸10mg/L、色氨酸10mg/L、酪氨酸10mg/L、缬氨酸10mg/L、氯化钙10mg/L、硫酸镁10mg/L、氯化钾10mg/L、磷酸二氢钾10mg/L、磷酸氢二钠10mg/L、碳酸氢钠0.1g/L、氢氧化钠0.1g/L、七水合硫酸亚铁10mg/L、六水合氯化镁10mg/L、硫酸铜10mg/L、丙酮酸钠10mg/L、二氯化锰10mg/L、亚硒酸钠10mg/L、钼酸铵0.001mg/L、丁酸钠10mg/L、丁二酸钠10mg/L、柠檬酸铁10mg/L、柠檬酸10mg/L、氯化钠1g/L、生物素1mg/L、氯化胆碱1mg/L、酒石酸胆碱1mg/L、肌醇1mg/L、尼克酰胺1mg/L、D-泛酸半钙1mg/L、吡哆醛1mg/L、硫胺素1mg/L、核黄素1mg/L、维生素B2 1mg/L、维生素D2 1mg/L、抗坏血酸1mg/L、维生素 K2 1mg/L、维生素K3 1mg/L、维生素B-12 0.1mg/L、对氨基苯甲酸1mg/L、叶酸1mg/L、D- 葡萄糖1000mg/L、胸苷10mg/L、谷胱苷肽还原型1mg/L、腐胺1mg/L、乙醇胺1mg/L、β- 巯基乙醇1mg/L、次黄嘌呤1mg/L、腺嘌呤1mg/L、盐酸鸟嘌呤1mg/L、核糖1mg/L、尿嘧啶 1mg/L、硫辛酸1mg/L、亚油酸1mL/L、胆固醇1mg/L、氨基多糖1mg/L、hepes 1g/L、胰岛素0.00001mg/L、氢化可的松0.00001mg/L、地塞米松0.00001mg/L、雌二醇孕酮0.00001mg/L、睾酮0.00001mg/L、酵母水解物10mg/l、水解乳蛋白10mg/l、酪蛋白水解物10mg/l。
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸100mg/L、精氨酸1100mg/L、天冬氨酸100mg/L、天冬酰氨100mg/L、胱氨酸100mg/L、半胱氨酸100mg/L、谷氨酰胺 1000mg/L、谷氨酸100mg/L、甘氨酸100mg/L、组氨酸100mg/L、羟脯氨酸100mg/L、异亮氨酸100mg/L、亮氨酸100mg/L、赖氨酸100mg/L、蛋氨酸100mg/L、苯丙氨酸1100mg/L、脯氨酸100mg/L、丝氨酸100mg/L、苏氨酸100mg/L、色氨酸100mg/L、酪氨酸100mg/L、缬氨酸100mg/L、氯化钙100mg/L、硫酸镁100mg/L、氯化钾1000mg/L、磷酸二氢钾100mg/L、磷酸氢二钠100mg/L、碳酸氢钠5g/L、氢氧化钠5g/L、七水合硫酸亚铁100mg/L、六水合氯化镁100mg/L、硫酸铜100mg/L、丙酮酸钠100mg/L、二氯化锰100mg/L、亚硒酸钠100mg/L、钼酸铵100mg/L、丁酸钠100mg/L、丁二酸钠100mg/L、柠檬酸铁100mg/L、柠檬酸100mg/L、氯化钠10g/L、生物素100mg/L、氯化胆碱100mg/L、酒石酸胆碱100mg/L、肌醇100mg/L、尼克酰胺100mg/L、D-泛酸半钙100mg/L、吡哆醛100mg/L、硫胺素100mg/L、核黄素100mg/L、维生素B2 100mg/L、维生素D2 100mg/L、抗坏血酸100mg/L、维生素K2 100mg/L、维生素 K3 100mg/L、维生素B-12100mg/L、对氨基苯甲酸100mg/L、叶酸100mg/L、D-葡萄糖 6000mg/L、胸苷100mg/L、谷胱苷肽还原型100mg/L、腐胺100mg/L、乙醇胺100mg/L、β- 巯基乙醇100mg/L、次黄嘌呤100mg/L、腺嘌呤100mg/L、盐酸鸟嘌呤100mg/L、核糖100mg/L、尿嘧啶100mg/L、硫辛酸100mg/L、亚油酸100mL/L、胆固醇100mg/L、氨基多糖100mg/L、 hepes 10g/L、胰岛素10mg/L、氢化可的松10mg/L、地塞米松10mg/L、雌二醇孕酮10mg/L、睾酮10mg/L、酵母水解物1000mg/l、水解乳蛋白1000mg/l、酪蛋白水解物1000mg/l。
本发明与现有技术相比，具有如下的优点和有益效果：
本发明一种杂交瘤细胞无血清培养基，完全不含有血清成份；生产的抗体纯度大幅度提高；该培养基同时具有高扩增速度及高抗体表达能力；杂交瘤细胞不需要驯化，可以直接从含血清培养直接过渡到无血清培养基培养；该培养基能长时间保持细胞活性，杂交瘤细胞培养时间大大延长，可以大大延长生产时间，提高抗体产量。
附图说明
此处所说明的附图用来提供对本发明实施例的进一步理解，构成本申请的一部分，并不构成对本发明实施例的限定。在附图中：
图1为分别采用本发明的培养基与DMEM/F12培养基培养杂交细胞(PHSA)生长曲线对比图；
图2为分别采用本发明的培养基与DMEM/F12培养基培养杂交细胞(PHSA)抗体表达对比图
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白，下面结合实施例，对本发明作进一步的详细说明，本发明的示意性实施方式及其说明仅用于解释本发明，并不作为对本发明的限定。
实施例1
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸50mg/L、精氨酸40mg/L、天冬氨酸30mg/L、天冬酰氨50mg/L、胱氨酸40mg/L、半胱氨酸70mg/L、谷氨酰胺600mg/L、谷氨酸20mg/L、甘氨酸30mg/L、组氨酸50mg/L、羟脯氨酸70mg/L、异亮氨酸80mg/L、亮氨酸30mg/L、赖氨酸90mg/L、蛋氨酸60mg/L、苯丙氨酸50mg/L、脯氨酸40mg/L、丝氨酸50mg/L、苏氨酸40mg/L、色氨酸30mg/L、酪氨酸50mg/L、缬氨酸30mg/L、氯化钙40mg/L、硫酸镁35mg/L、氯化钾700mg/L、磷酸二氢钾60mg/L、磷酸氢二钠40mg/L、碳酸氢钠1g/L、氢氧化钠3g/L、七水合硫酸亚铁40mg/L、六水合氯化镁70mg/L、硫酸铜30mg/L、丙酮酸钠 40mg/L、二氯化锰50mg/L、亚硒酸钠40mg/L、钼酸铵5mg/L、丁酸钠70mg/L、丁二酸钠30mg/L、柠檬酸铁20mg/L、柠檬酸55mg/L、氯化钠3g/L、生物素40mg/L、氯化胆碱25mg/L、酒石酸胆碱60mg/L、肌醇40mg/L、尼克酰胺35mg/L、D-泛酸半钙40mg/L、吡哆醛10mg/L、硫胺素100mg/L、核黄素70mg/L、维生素B2 50mg/L、维生素D2 20mg/L、抗坏血酸60mg/L、维生素K2 50mg/L、维生素K3 20mg/L、维生素B-12 10mg/L、对氨基苯甲酸15mg/L、叶酸 60mg/L、D-葡萄糖4000mg/L、胸苷90mg/L、谷胱苷肽还原型30mg/L、腐胺50mg/L、乙醇胺45mg/L、β-巯基乙醇80mg/L、次黄嘌呤70mg/L、腺嘌呤45mg/L、盐酸鸟嘌呤50mg/L、核糖45mg/L、尿嘧啶55mg/L、硫辛酸60mg/L、亚油酸30mL/L、胆固醇80mg/L、氨基多糖 60mg/L、hepes 5g/L、胰岛素2mg/L、氢化可的松40mg/L、地塞米松0.5mg/L、雌二醇孕酮 1mg/L、睾酮1mg/L、酵母水解物600mg/l、水解乳蛋白800mg/l、酪蛋白水解物600mg/l。
实施例2
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸10mg/L、精氨酸10mg/L、天冬氨酸10mg/L、天冬酰氨10mg/L、胱氨酸10mg/L、半胱氨酸10mg/L、谷氨酰胺100mg/L、谷氨酸10mg/L、甘氨酸10mg/L、组氨酸10mg/L、羟脯氨酸10mg/L、异亮氨酸10mg/L、亮氨酸10mg/L、赖氨酸10mg/L、蛋氨酸10mg/L、苯丙氨酸10mg/L、脯氨酸10mg/L、丝氨酸10mg/L、苏氨酸10mg/L、色氨酸10mg/L、酪氨酸10mg/L、缬氨酸10mg/L、氯化钙10mg/L、硫酸镁10mg/L、氯化钾10mg/L、磷酸二氢钾10mg/L、磷酸氢二钠10mg/L、碳酸氢钠0.1g/L、氢氧化钠0.1g/L、七水合硫酸亚铁10mg/L、六水合氯化镁10mg/L、硫酸铜10mg/L、丙酮酸钠10mg/L、二氯化锰10mg/L、亚硒酸钠10mg/L、钼酸铵0.001mg/L、丁酸钠10mg/L、丁二酸钠10mg/L、柠檬酸铁10mg/L、柠檬酸10mg/L、氯化钠1g/L、生物素1mg/L、氯化胆碱1mg/L、酒石酸胆碱1mg/L、肌醇1mg/L、尼克酰胺1mg/L、D-泛酸半钙1mg/L、吡哆醛1mg/L、硫胺素1mg/L、核黄素1mg/L、维生素B2 1mg/L、维生素D2 1mg/L、抗坏血酸1mg/L、维生素 K2 1mg/L、维生素K3 1mg/L、维生素B-12 0.1mg/L、对氨基苯甲酸1mg/L、叶酸1mg/L、D- 葡萄糖1000mg/L、胸苷10mg/L、谷胱苷肽还原型1mg/L、腐胺1mg/L、乙醇胺1mg/L、β- 巯基乙醇1mg/L、次黄嘌呤1mg/L、腺嘌呤1mg/L、盐酸鸟嘌呤1mg/L、核糖1mg/L、尿嘧啶 1mg/L、硫辛酸1mg/L、亚油酸1mL/L、胆固醇1mg/L、氨基多糖1mg/L、hepes 1g/L、胰岛素0.00001mg/L、氢化可的松0.00001mg/L、地塞米松0.00001mg/L、雌二醇孕酮0.00001mg/L、睾酮0.00001mg/L、酵母水解物10mg/l、水解乳蛋白10mg/l、酪蛋白水解物10mg/l。
实施例3
一种杂交瘤细胞无血清培养基，包括如下浓度的原料：丙氨酸100mg/L、精氨酸1100mg/L、天冬氨酸100mg/L、天冬酰氨100mg/L、胱氨酸100mg/L、半胱氨酸100mg/L、谷氨酰胺 1000mg/L、谷氨酸100mg/L、甘氨酸100mg/L、组氨酸100mg/L、羟脯氨酸100mg/L、异亮氨酸100mg/L、亮氨酸100mg/L、赖氨酸100mg/L、蛋氨酸100mg/L、苯丙氨酸1100mg/L、脯氨酸100mg/L、丝氨酸100mg/L、苏氨酸100mg/L、色氨酸100mg/L、酪氨酸100mg/L、缬氨酸100mg/L、氯化钙100mg/L、硫酸镁100mg/L、氯化钾1000mg/L、磷酸二氢钾100mg/L、磷酸氢二钠100mg/L、碳酸氢钠5g/L、氢氧化钠5g/L、七水合硫酸亚铁100mg/L、六水合氯化镁100mg/L、硫酸铜100mg/L、丙酮酸钠100mg/L、二氯化锰100mg/L、亚硒酸钠100mg/L、钼酸铵100mg/L、丁酸钠100mg/L、丁二酸钠100mg/L、柠檬酸铁100mg/L、柠檬酸100mg/L、氯化钠10g/L、生物素100mg/L、氯化胆碱100mg/L、酒石酸胆碱100mg/L、肌醇100mg/L、尼克酰胺100mg/L、D-泛酸半钙100mg/L、吡哆醛100mg/L、硫胺素100mg/L、核黄素100mg/L、维生素B2 100mg/L、维生素D2 100mg/L、抗坏血酸100mg/L、维生素K2 100mg/L、维生素 K3 100mg/L、维生素B-12100mg/L、对氨基苯甲酸100mg/L、叶酸100mg/L、D-葡萄糖 6000mg/L、胸苷100mg/L、谷胱苷肽还原型100mg/L、腐胺100mg/L、乙醇胺100mg/L、β- 巯基乙醇100mg/L、次黄嘌呤100mg/L、腺嘌呤100mg/L、盐酸鸟嘌呤100mg/L、核糖100mg/L、尿嘧啶100mg/L、硫辛酸100mg/L、亚油酸100mL/L、胆固醇100mg/L、氨基多糖100mg/L、 hepes 10g/L、胰岛素10mg/L、氢化可的松10mg/L、地塞米松10mg/L、雌二醇孕酮10mg/L、睾酮10mg/L、酵母水解物1000mg/l、水解乳蛋白1000mg/l、酪蛋白水解物1000mg/l。
实施例4
实验组：采用本发明实施例1的细胞培养基；
对照组：采用DMEM/F12+10％血清的培养基。
用上述实验组与对比组的培养基培养杂交瘤细胞。
如图1和图2中，小的菱形方块代表实验组，大的正方形块代表对照组。由图1可以看出在实验组的培养基中能长期培养杂交瘤细胞，而对照组用血清培养杂交瘤细胞，细胞增值到6-7天达到高峰后，细胞开始大量死亡。由图2可以看出在实验组的培养基培养杂交瘤细胞能长期持续获得抗体，且抗体产量明显高于对照组。
以上所述的具体实施方式，对本发明的目的、技术方案和有益效果进行了进一步详细说明，所应理解的是，以上所述仅为本发明的具体实施方式而已，并不用于限定本发明的保护范围，凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (5)

1.一种杂交瘤细胞无血清培养基，其特征在于，包括如下浓度的原料：丙氨酸10-100mg/L、精氨酸10-100mg/L、天冬氨酸10-100mg/L、天冬酰氨10-100mg/L、胱氨酸10-100mg/L、半胱氨酸10-100mg/L、谷氨酰胺100-1000mg/L、谷氨酸10-100mg/L、甘氨酸10-100mg/L、组氨酸10-100mg/L、羟脯氨酸10-100mg/L、异亮氨酸10-100mg/L、亮氨酸10-100mg/L、赖氨酸10-100mg/L、蛋氨酸10-100mg/L、苯丙氨酸10-100mg/L、脯氨酸10-100mg/L、丝氨酸10-100mg/L、苏氨酸10-100mg/L、色氨酸10-100mg/L、酪氨酸10-100mg/L、缬氨酸10-100mg/L、氯化钙10-100mg/L、硫酸镁10-100mg/L、氯化钾10-1000mg/L、磷酸二氢钾10-100mg/L、磷酸氢二钠10-100mg/L、碳酸氢钠0.1-5g/L、氢氧化钠0.1-5g/L、七水合硫酸亚铁10-100mg/L、六水合氯化镁10-100mg/L、硫酸铜10-100mg/L、丙酮酸钠10-100mg/L、二氯化锰10-100mg/L、亚硒酸钠10-100mg/L、钼酸铵0.001-100mg/L、丁酸钠10-100mg/L、丁二酸钠10-100mg/L、柠檬酸铁10-100mg/L、柠檬酸10-100mg/L、氯化钠1-10g/L、生物素1-100mg/L、氯化胆碱1-100mg/L、酒石酸胆碱1-100mg/L、肌醇1-100mg/L、尼克酰胺1-100mg/L、D-泛酸半钙1-100mg/L、吡哆醛1-100mg/L、硫胺素1-100mg/L、核黄素1-100mg/L、维生素B2 1-100mg/L、维生素D2 1-100mg/L、抗坏血酸1-100mg/L、维生素K2 1-100mg/L、维生素K3 1-100mg/L、维生素B-12 0.1-100mg/L、对氨基苯甲酸1-100mg/L、叶酸1-100mg/L、D-葡萄糖1000-6000mg/L、胸苷10-100mg/L、谷胱苷肽还原型1-100mg/L、腐胺1-100mg/L、乙醇胺1-100mg/L、β-巯基乙醇1-100mg/L、次黄嘌呤1-100mg/L、腺嘌呤1-100mg/L、盐酸鸟嘌呤1-100mg/L、核糖1-100mg/L、尿嘧啶1-100mg/L、硫辛酸1-100mg/L、亚油酸1-100mL/L、胆固醇1-100mg/L、氨基多糖1-100mg/L、hepes 1-10g/L、胰岛素0.00001-10mg/L、氢化可的松0.00001-10mg/L、地塞米松0.00001-10mg/L、雌二醇孕酮0.00001-10mg/L、睾酮0.00001-10mg/L、酵母水解物10-1000mg/l、水解乳蛋白10-1000mg/l、酪蛋白水解物10-1000mg/l。
2.根据权利要求1所述的一种杂交瘤细胞无血清培养基，其特征在于，包括如下浓度的原料：丙氨酸30-100mg/L、精氨酸10-80mg/L、天冬氨酸10-70mg/L、天冬酰氨20-100mg/L、胱氨酸10-90mg/L、半胱氨酸50-100mg/L、谷氨酰胺300-1000mg/L、谷氨酸10-70mg/L、甘氨酸20-80mg/L、组氨酸10-60mg/L、羟脯氨酸40-100mg/L、异亮氨酸20-100mg/L、亮氨酸10-60mg/L、赖氨酸50-100mg/L、蛋氨酸30-90mg/L、苯丙氨酸40-100mg/L、脯氨酸20-60mg/L、丝氨酸20-60mg/L、苏氨酸20-60mg/L、色氨酸20-60mg/L、酪氨酸20-60mg/L、缬氨酸20-60mg/L、氯化钙20-60mg/L、硫酸镁20-60mg/L、氯化钾300-1000mg/L、磷酸二氢钾30-70mg/L、磷酸氢二钠30-70mg/L、碳酸氢钠0.1-2g/L、氢氧化钠2-5g/L、七水合硫酸亚铁30-70mg/L、六水合氯化镁30-70mg/L、硫酸铜10-90mg/L、丙酮酸钠30-70mg/L、二氯化锰40-100mg/L、亚硒酸钠30-70mg/L、钼酸铵10-100mg/L、丁酸钠30-70mg/L、丁二酸钠30-70mg/L、柠檬酸铁10-50mg/L、柠檬酸30-70mg/L、氯化钠1-6g/L、生物素30-70mg/L、氯化胆碱20-70mg/L、酒石酸胆碱30-60mg/L、肌醇40-80mg/L、尼克酰胺30-70mg/L、D-泛酸半钙30-50mg/L、吡哆醛10-70mg/L、硫胺素30-100mg/L、核黄素1-80mg/L、维生素B230-70mg/L、维生素D2 20-70mg/L、抗坏血酸30-80mg/L、维生素K2 10-70mg/L、维生素K310-60mg/L、维生素B-12 0.1-50mg/L、对氨基苯甲酸10-40mg/L、叶酸30-70mg/L、D-葡萄糖3000-6000mg/L、胸苷80-100mg/L、谷胱苷肽还原型10-50mg/L、腐胺30-70mg/L、乙醇胺30-70mg/L、β-巯基乙醇40-100mg/L、次黄嘌呤30-90mg/L、腺嘌呤30-70mg/L、盐酸鸟嘌呤30-70mg/L、核糖30-70mg/L、尿嘧啶20-100mg/L、硫辛酸30-80mg/L、亚油酸15-60mL/L、胆固醇70-100mg/L、氨基多糖50-90mg/L、hepes 3-8g/L、胰岛素0.00001-5mg/L、氢化可的松1-10mg/L、地塞米松0.00001-2mg/L、雌二醇孕酮0.00001-3mg/L、睾酮0.5-2mg/L、酵母水解物400-1000mg/l、水解乳蛋白500-1000mg/l、酪蛋白水解物10-1000mg/l。
3.根据权利要求1所述的一种杂交瘤细胞无血清培养基，其特征在于，包括如下浓度的原料：丙氨酸50mg/L、精氨酸40mg/L、天冬氨酸30mg/L、天冬酰氨50mg/L、胱氨酸40mg/L、半胱氨酸70mg/L、谷氨酰胺600mg/L、谷氨酸20mg/L、甘氨酸30mg/L、组氨酸50mg/L、羟脯氨酸70mg/L、异亮氨酸80mg/L、亮氨酸30mg/L、赖氨酸90mg/L、蛋氨酸60mg/L、苯丙氨酸50mg/L、脯氨酸40mg/L、丝氨酸50mg/L、苏氨酸40mg/L、色氨酸30mg/L、酪氨酸50mg/L、缬氨酸30mg/L、氯化钙40mg/L、硫酸镁35mg/L、氯化钾700mg/L、磷酸二氢钾60mg/L、磷酸氢二钠40mg/L、碳酸氢钠1g/L、氢氧化钠3g/L、七水合硫酸亚铁40mg/L、六水合氯化镁70mg/L、硫酸铜30mg/L、丙酮酸钠40mg/L、二氯化锰50mg/L、亚硒酸钠40mg/L、钼酸铵5mg/L、丁酸钠70mg/L、丁二酸钠30mg/L、柠檬酸铁20mg/L、柠檬酸55mg/L、氯化钠3g/L、生物素40mg/L、氯化胆碱25mg/L、酒石酸胆碱60mg/L、肌醇40mg/L、尼克酰胺35mg/L、D-泛酸半钙40mg/L、吡哆醛10mg/L、硫胺素100mg/L、核黄素70mg/L、维生素B2 50mg/L、维生素D2 20mg/L、抗坏血酸60mg/L、维生素K2 50mg/L、维生素K3 20mg/L、维生素B-1210mg/L、对氨基苯甲酸15mg/L、叶酸60mg/L、D-葡萄糖4000mg/L、胸苷90mg/L、谷胱苷肽还原型30mg/L、腐胺50mg/L、乙醇胺45mg/L、β-巯基乙醇80mg/L、次黄嘌呤70mg/L、腺嘌呤45mg/L、盐酸鸟嘌呤50mg/L、核糖45mg/L、尿嘧啶55mg/L、硫辛酸60mg/L、亚油酸30mL/L、胆固醇80mg/L、氨基多糖60mg/L、hepes 5g/L、胰岛素2mg/L、氢化可的松40mg/L、地塞米松0.5mg/L、雌二醇孕酮1mg/L、睾酮1mg/L、酵母水解物600mg/l、水解乳蛋白800mg/l、酪蛋白水解物600mg/l。
4.根据权利要求1所述的一种杂交瘤细胞无血清培养基，其特征在于，包括如下浓度的原料：丙氨酸10mg/L、精氨酸10mg/L、天冬氨酸10mg/L、天冬酰氨10mg/L、胱氨酸10mg/L、半胱氨酸10mg/L、谷氨酰胺100mg/L、谷氨酸10mg/L、甘氨酸10mg/L、组氨酸10mg/L、羟脯氨酸10mg/L、异亮氨酸10mg/L、亮氨酸10mg/L、赖氨酸10mg/L、蛋氨酸10mg/L、苯丙氨酸10mg/L、脯氨酸10mg/L、丝氨酸10mg/L、苏氨酸10mg/L、色氨酸10mg/L、酪氨酸10mg/L、缬氨酸10mg/L、氯化钙10mg/L、硫酸镁10mg/L、氯化钾10mg/L、磷酸二氢钾10mg/L、磷酸氢二钠10mg/L、碳酸氢钠0.1g/L、氢氧化钠0.1g/L、七水合硫酸亚铁10mg/L、六水合氯化镁10mg/L、硫酸铜10mg/L、丙酮酸钠10mg/L、二氯化锰10mg/L、亚硒酸钠10mg/L、钼酸铵0.001mg/L、丁酸钠10mg/L、丁二酸钠10mg/L、柠檬酸铁10mg/L、柠檬酸10mg/L、氯化钠1g/L、生物素1mg/L、氯化胆碱1mg/L、酒石酸胆碱1mg/L、肌醇1mg/L、尼克酰胺1mg/L、D-泛酸半钙1mg/L、吡哆醛1mg/L、硫胺素1mg/L、核黄素1mg/L、维生素B2 1mg/L、维生素D2 1mg/L、抗坏血酸1mg/L、维生素K2 1mg/L、维生素K3 1mg/L、维生素B-12 0.1mg/L、对氨基苯甲酸1mg/L、叶酸1mg/L、D-葡萄糖1000mg/L、胸苷10mg/L、谷胱苷肽还原型1mg/L、腐胺1mg/L、乙醇胺1mg/L、β-巯基乙醇1mg/L、次黄嘌呤1mg/L、腺嘌呤1mg/L、盐酸鸟嘌呤1mg/L、核糖1mg/L、尿嘧啶1mg/L、硫辛酸1mg/L、亚油酸1mL/L、胆固醇1mg/L、氨基多糖1mg/L、hepes 1g/L、胰岛素0.00001mg/L、氢化可的松0.00001mg/L、地塞米松0.00001mg/L、雌二醇孕酮0.00001mg/L、睾酮0.00001mg/L、酵母水解物10mg/l、水解乳蛋白10mg/l、酪蛋白水解物10mg/l。
5.根据权利要求1所述的一种杂交瘤细胞无血清培养基，其特征在于，包括如下浓度的原料：丙氨酸100mg/L、精氨酸1100mg/L、天冬氨酸100mg/L、天冬酰氨100mg/L、胱氨酸100mg/L、半胱氨酸100mg/L、谷氨酰胺1000mg/L、谷氨酸100mg/L、甘氨酸100mg/L、组氨酸100mg/L、羟脯氨酸100mg/L、异亮氨酸100mg/L、亮氨酸100mg/L、赖氨酸100mg/L、蛋氨酸100mg/L、苯丙氨酸1100mg/L、脯氨酸100mg/L、丝氨酸100mg/L、苏氨酸100mg/L、色氨酸100mg/L、酪氨酸100mg/L、缬氨酸100mg/L、氯化钙100mg/L、硫酸镁100mg/L、氯化钾1000mg/L、磷酸二氢钾100mg/L、磷酸氢二钠100mg/L、碳酸氢钠5g/L、氢氧化钠5g/L、七水合硫酸亚铁100mg/L、六水合氯化镁100mg/L、硫酸铜100mg/L、丙酮酸钠100mg/L、二氯化锰100mg/L、亚硒酸钠100mg/L、钼酸铵100mg/L、丁酸钠100mg/L、丁二酸钠100mg/L、柠檬酸铁100mg/L、柠檬酸100mg/L、氯化钠10g/L、生物素100mg/L、氯化胆碱100mg/L、酒石酸胆碱100mg/L、肌醇100mg/L、尼克酰胺100mg/L、D-泛酸半钙100mg/L、吡哆醛100mg/L、硫胺素100mg/L、核黄素100mg/L、维生素B2 100mg/L、维生素D2 100mg/L、抗坏血酸100mg/L、维生素K2 100mg/L、维生素K3 100mg/L、维生素B-12 100mg/L、对氨基苯甲酸100mg/L、叶酸100mg/L、D-葡萄糖6000mg/L、胸苷100mg/L、谷胱苷肽还原型100mg/L、腐胺100mg/L、乙醇胺100mg/L、β-巯基乙醇100mg/L、次黄嘌呤100mg/L、腺嘌呤100mg/L、盐酸鸟嘌呤100mg/L、核糖100mg/L、尿嘧啶100mg/L、硫辛酸100mg/L、亚油酸100mL/L、胆固醇100mg/L、氨基多糖100mg/L、hepes 10g/L、胰岛素10mg/L、氢化可的松10mg/L、地塞米松10mg/L、雌二醇孕酮10mg/L、睾酮10mg/L、酵母水解物1000mg/l、水解乳蛋白1000mg/l、酪蛋白水解物1000mg/l。
CN201711319522.8A 2017-12-12 2017-12-12 一种杂交瘤细胞无血清培养基 Withdrawn CN107881142A (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711319522.8A CN107881142A (zh) 2017-12-12 2017-12-12 一种杂交瘤细胞无血清培养基

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711319522.8A CN107881142A (zh) 2017-12-12 2017-12-12 一种杂交瘤细胞无血清培养基

Publications (1)

Publication Number Publication Date
CN107881142A true CN107881142A (zh) 2018-04-06

Family

ID=61774069

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711319522.8A Withdrawn CN107881142A (zh) 2017-12-12 2017-12-12 一种杂交瘤细胞无血清培养基

Country Status (1)

Country Link
CN (1) CN107881142A (zh)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108546684A (zh) * 2018-04-28 2018-09-18 深圳市菲鹏生物制药股份有限公司 一种杂交瘤细胞培养基及其制备方法
CN110117573A (zh) * 2019-04-15 2019-08-13 河北省科学院生物研究所 一种无血清细胞培养基及其应用
CN114480294A (zh) * 2021-12-15 2022-05-13 上海捷门生物技术有限公司 一种适合杂交瘤细胞贴壁生长的无血清培养基
CN115612658A (zh) * 2022-12-19 2023-01-17 天信和(苏州)生物科技有限公司 一种st细胞悬浮驯化的培养基及其应用

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108546684A (zh) * 2018-04-28 2018-09-18 深圳市菲鹏生物制药股份有限公司 一种杂交瘤细胞培养基及其制备方法
CN110117573A (zh) * 2019-04-15 2019-08-13 河北省科学院生物研究所 一种无血清细胞培养基及其应用
CN114480294A (zh) * 2021-12-15 2022-05-13 上海捷门生物技术有限公司 一种适合杂交瘤细胞贴壁生长的无血清培养基
CN114480294B (zh) * 2021-12-15 2024-01-26 上海捷门生物技术有限公司 一种适合杂交瘤细胞贴壁生长的无血清培养基
CN115612658A (zh) * 2022-12-19 2023-01-17 天信和(苏州)生物科技有限公司 一种st细胞悬浮驯化的培养基及其应用

Similar Documents

Publication Publication Date Title
CN107881142A (zh) 一种杂交瘤细胞无血清培养基
CN107988146A (zh) Cho细胞无蛋白培养基的制备方法
CN107881143A (zh) 一种cho细胞无血清培养基
CN101760442A (zh) 适于mdck细胞大规模贴壁培养和单细胞悬浮培养的无血清培养基
CN105018416A (zh) 一种悬浮培养bhk-21细胞的无血清无动物源培养基及其配制方法
CN110628697A (zh) 一种用于vero无血清细胞培养及相应病毒生产的无血清培养基
CN101974481A (zh) 一种用于多种肾脏组织来源细胞生长的无血清培养基
CN100569940C (zh) 人胚胎肾(hek)293细胞无血清培养基
KR20070058584A (ko) 인간 배아 줄기 세포의 배양
CN106635953B (zh) 无血清无蛋白细胞培养基
CN104278009B (zh) 无血清培养基及其用途和一种猪瘟病毒的培养方法
CN104328158B (zh) 适于动物细胞表达产品大规模生产的化学成分明确培养基
CN110894487B (zh) 一种无血清无蛋白cho细胞培养基及其制备方法、应用
CN104357379A (zh) 干细胞培养基
CN107201344A (zh) 一种动物细胞融合基础培养基
CN106754634A (zh) 一种无血清培养基及其制备方法
CN107236708A (zh) 一种支持HeLa细胞贴壁培养的无血清培养基
CN110643568A (zh) 一种用于bhk-21细胞培养及相应病毒生产的低血清培养基
CN105087461A (zh) 一种pk细胞培养基
CN108060128A (zh) 免疫细胞用基础培养基的生产方法
CN104630132B (zh) 一种培养Marc‑145细胞用的培养基及其制备方法
CN108060115A (zh) 一种cho细胞无蛋白培养基
CN105087460A (zh) 一种st细胞培养基
AU2020331685A1 (en) Cell culture medium for cultivating cells, method for cultivating cells, and method for expressing at least one recombinant protein in a cell culture
CN110117573A (zh) 一种无血清细胞培养基及其应用

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
WW01 Invention patent application withdrawn after publication

Application publication date: 20180406

WW01 Invention patent application withdrawn after publication