The first recombinant human coagulation factor VIII of human origin: human cell line and manufacturing characteristics.
The first recombinant human coagulation factor VIII of human origin: human cell line and manufacturing characteristics.
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DOI：
10.1111/j.1600-0609.2012.01804.x
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发表时间：
2012-08
影响因子：
3.1
通讯作者：
Schröder C
中科院分区：
文献类型：
--
作者：
Casademunt E;Martinelle K;Jernberg M;Winge S;Tiemeyer M;Biesert L;Knaub S;Walter O;Schröder C
自20世纪90年代初以来，在仓鼠细胞中生产的重组人凝血因子VIII（rhFVIII）已可用于血友病A治疗。然而，这些蛋白质的翻译后修饰与天然人FVIII的翻译后修饰不同，这可能导致免疫原性反应和rhFVIII抑制剂的产生。这是第一次，在人类宿主细胞系中产生的rhFVIII是可用的。我们在此描述了rhFVIII的第一个人生产细胞系的建立和这种新产品的生产工艺。表达rhFVIII的人细胞系来源于用FVIII表达质粒转染的人胚肾（HEK）293 F细胞。经过广泛检测，未检测到病毒或病毒样颗粒。严格控制的生产过程完全不含动物或人源性添加材料。采用过滤和色谱步骤组合的多步纯化确保了杂质的有效去除。溶剂/去污剂处理和20 nm孔径纳滤步骤首次用于rhFVIII生产，可有效消除任何假设存在的病毒。与仓鼠细胞衍生产品相比，该rhFVIII产品不含仓鼠样表位，预期其可能具有免疫原性。HEK 293 F细胞，其亲本细胞系HEK 293已被研究人员使用了几十年，是rhFVIII的合适生产细胞系，将有助于避免免疫原性表位。我们开发了现代化的生产工艺，以确保最高水平的纯度和病原体安全性。
Since the early 1990s, recombinant human clotting factor VIII (rhFVIII) produced in hamster cells has been available for haemophilia A treatment. However, the post-translational modifications of these proteins are not identical to those of native human FVIII, which may lead to immunogenic reactions and the development of inhibitors against rhFVIII. For the first time, rhFVIII produced in a human host cell line is available. We describe here the establishment of the first human production cell line for rhFVIII and the manufacturing process of this novel product. A human cell line expressing rhFVIII was derived from human embryonic kidney (HEK) 293 F cells transfected with an FVIII expression plasmid. No virus or virus-like particles could be detected following extensive testing. The stringently controlled production process is completely free from added materials of animal or human origin. Multistep purification employing a combination of filtration and chromatography steps ensures the efficient removal of impurities. Solvent/detergent treatment and a 20 nm pore size nanofiltration step, used for the first time in rhFVIII manufacturing, efficiently eliminate any hypothetically present viruses. In contrast to hamster cell-derived products, this rhFVIII product does not contain hamster-like epitopes, which might be expected to be immunogenic. HEK 293 F cells, whose parental cell line HEK 293 has been used by researchers for decades, are a suitable production cell line for rhFVIII and will help avoid immunogenic epitopes. A modern manufacturing process has been developed to ensure the highest level of purity and pathogen safety.
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影响因子：
2.9
作者：
Dichtelmueller, Herbert O.;Biesert, Lothar;Poelsler, Gerhard
通讯作者：
Poelsler, Gerhard
影响因子：
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作者：
GRAHAM, FL;SMILEY, J;NAIRN, R
通讯作者：
NAIRN, R
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作者：
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影响因子：
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作者：
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通讯作者：
Uren, E
影响因子：
14.9
作者：
Durocher, Y;Perret, S;Kamen, A
通讯作者：
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