CN107881143A - 一种cho细胞无血清培养基 - Google Patents

一种cho细胞无血清培养基 Download PDF

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CN107881143A
CN107881143A CN201711321218.7A CN201711321218A CN107881143A CN 107881143 A CN107881143 A CN 107881143A CN 201711321218 A CN201711321218 A CN 201711321218A CN 107881143 A CN107881143 A CN 107881143A
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刘才明
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Chengdu Source Biotechnology Co Ltd
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Abstract

本发明公开了一种CHO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、酵母水解物、乳蛋白水解物、酪蛋白水解物。本发明培养基，完全不含有血清成份，有利于蛋白的纯化工艺；CHO细胞无需通过减血清的适应过程；能长期保持细胞活性，延长了蛋白表达生产时间，大幅度降低了无血清培养基的成本，为企业降低生产成本。

Description

一种CHO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CHO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。世界上有一半的重组药物表达用的是哺乳动物细胞，而这其中70％是中国仓鼠卵巢细胞(CHO细胞)表达。可见CHO是重组药物生产最重要的宿主细胞。而在培养CHO细胞生产药物的工艺上，多数采用有血清和蛋白成份的添加，而这些成份的添加往往可能带来外源的污染或者造成目标蛋白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现有技术CHO细胞培养基中均含有血清，但是血清成份的添加带来外源的污染或者造成目标蛋白在分离纯化上难度的增加，目的在于提供一种CHO细胞无血清培养基，完全不含有血清成份，有利于蛋白的纯化工艺；该培养基在CHO培养中， CHO细胞无需通过减血清的适应过程；能长期保持细胞活性，延长了蛋白表达生产时间，大幅度降低了无血清培养基的成本，为企业降低生产成本。
本发明通过下述技术方案实现：
一种CHO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、丁酸钠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K31-100mg/L、维生素E1-100mg/l、维生素C 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、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、酵母水解物10-1000mg/l、乳蛋白水解物10-1000mg/l、酪蛋白水解物10-1000mg/l。
一种CHO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、丁酸钠 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E 10-80mg/l、维生素C 30-100mg/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、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、酵母水解物300-1000mg/l、乳蛋白水解物400-1000mg/l、酪蛋白水解物 500-1000mg/l。
一种CHO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、丁酸钠70mg/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E 50mg/l、维生素C 60mg/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、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、酵母水解物600mg/l、乳蛋白水解物500mg/l、酪蛋白水解物300mg/l。
一种CHO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、丁酸钠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E 10mg/l、维生素C 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、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、酵母水解物10mg/l、乳蛋白水解物10mg/l、酪蛋白水解物10mg/l。
一种CHO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、丁酸钠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E 100mg/l、维生素C 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、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、酵母水解物1000mg/l、乳蛋白水解物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CHO细胞无血清培养基，完全不含有血清成份，有利于蛋白的纯化工艺；该培养基在CHO培养中，CHO细胞无需通过减血清的适应过程；能长期保持细胞活性，延长了蛋白表达生产时间，大幅度降低了无血清培养基的成本，为企业降低生产成本。
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CHO细胞生长曲线对比图；
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CHO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、丁酸钠70mg/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E 50mg/l、维生素C 60mg/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、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、酵母水解物600mg/l、乳蛋白水解物500mg/l、酪蛋白水解物300mg/l。
实施例2
一种CHO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、丁酸钠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E 10mg/l、维生素C 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、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、酵母水解物10mg/l、乳蛋白水解物10mg/l、酪蛋白水解物10mg/l。
实施例3
一种CHO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、丁酸钠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E 100mg/l、维生素C 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、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、酵母水解物1000mg/l、乳蛋白水解物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CHO细胞。
如图1中，小的菱形方块代表实验组，大的正方形块代表对照组，横坐标代表细胞培养天数，纵坐标代表细胞密度×106/mL。由图1可以看出CHO细胞在实验组的培养基中的生长数量与增殖速率明显高于对照组，本发明培养基能满足CHO细胞的生长，培养到三周(21 天)时细胞还存活状态，此时细胞大多贴壁状态，CHO再通过胰酶消化悬浮后还能继续培养和蛋白生产表达；而对照组细胞培养到第七天时细胞停止了增值，并开始死亡；因此本发明的培养基能有效避免细胞凋亡，可以大大延长CHO细胞的生产周期，从而大幅度节约生产成本。
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CHO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、丁酸钠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E 1-100mg/l、维生素C 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、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、酵母水解物10-1000mg/l、乳蛋白水解物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CHO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、丁酸钠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E 10-80mg/l、维生素C 30-100mg/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、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、酵母水解物300-1000mg/l、乳蛋白水解物400-1000mg/l、酪蛋白水解物500-1000mg/l。
3.根据权利要求1所述的一种CHO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、丁酸钠70mg/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E 50mg/l、维生素C 60mg/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、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、酵母水解物600mg/l、乳蛋白水解物500mg/l、酪蛋白水解物300mg/l。
4.根据权利要求1所述的一种CHO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、丁酸钠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D21mg/L、抗坏血酸1mg/L、维生素K2 1mg/L、维生素K3 1mg/L、维生素E 10mg/l、维生素C 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、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、酵母水解物10mg/l、乳蛋白水解物10mg/l、酪蛋白水解物10mg/l。
5.根据权利要求1所述的一种CHO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、丁酸钠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E 100mg/l、维生素C 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、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、酵母水解物1000mg/l、乳蛋白水解物1000mg/l、酪蛋白水解物1000mg/l。
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Cited By (4)

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Publication number Priority date Publication date Assignee Title
CN110343666A (zh) * 2019-07-10 2019-10-18 通化东宝生物科技有限公司 一种cho细胞培养的补料培养基及其制备方法和应用
CN110592000A (zh) * 2019-08-13 2019-12-20 苏州易迈吉生物医药科技有限公司 一种支持bhk细胞高密度悬浮培养的无血清培养基
CN115404201A (zh) * 2022-10-17 2022-11-29 湖南爱备康生物科技有限公司 一种提高干细胞外泌体分泌的无血清培养基
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
CN110343666A (zh) * 2019-07-10 2019-10-18 通化东宝生物科技有限公司 一种cho细胞培养的补料培养基及其制备方法和应用
CN110343666B (zh) * 2019-07-10 2023-05-30 通化东宝药业股份有限公司 一种cho细胞培养的补料培养基及其制备方法和应用
CN110592000A (zh) * 2019-08-13 2019-12-20 苏州易迈吉生物医药科技有限公司 一种支持bhk细胞高密度悬浮培养的无血清培养基
CN115404201A (zh) * 2022-10-17 2022-11-29 湖南爱备康生物科技有限公司 一种提高干细胞外泌体分泌的无血清培养基
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

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