A novel simple method to purify recombinant soluble human complement receptor type 1 (sCR1) from CHO cell culture
A novel simple method to purify recombinant soluble human complement receptor type 1 (sCR1) from CHO cell culture
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一种从 CHO 细胞培养物中纯化重组可溶性人补体受体 1 型 (sCR1) 的新颖简单方法
DOI：
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Pi
中科院分区：
文献类型：
--
作者：
Hisamune Kato;Takehiro Inoue;N. Ishii;Y. Murakami;M. Matsumura;T. Seya;Pi
人补体受体1型（CR 1，C3 B/C4 B受体）是一种在人红细胞、外周血白细胞、血浆和肾小球足细胞上表达的多态性膜糖蛋白，其由跨膜和胞质结构域组成，具有30个重复的同源蛋白结构域，称为短共有重复序列（SCR）。在过去的几年中，CR 1已被用作炎症和免疫系统的抑制剂。近年来，研究发现CR 1具有抑制异种移植超急性排斥反应的作用，可用于治疗自身免疫性疾病。CR 1的可溶性形式称为sCR 1，是通过在C-末端切割跨膜结构域而形成的重组CR 1，并且已在中国人卵巢（CHO）细胞中表达。目前已报道了几种从CHO细胞中纯化sCR 1的方法，但大多数方法步骤复杂，成本高。此外，这些方法大多在易于使sCR 1变性并导致sCR 1失去其活性的pH条件下进行。本文报道了一种从CHO细胞中快速、有效地纯化sCR 1的方法。新方法包括两个阶段的细胞培养，通过培养细胞在血清培养基，然后无血清培养基，和两个阶段的柱纯化，通过肝素和凝胶过滤柱色谱法。该方法简单、有效、收率高、纯度高。纯化的sCR 1在体内和体外均具有抑制补体激活的活性。
The human complement receptor type 1 (CR1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains knows as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently, it is reported that CR1 was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CR1, called sCR1, is a recombinant CR1 by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCR1 losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity. Furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activationin vivo andex vivo.
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可溶性 CR1 在补体和中性粒细胞介导的组织损伤中的保护作用。
DOI：
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发表时间：
1992
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
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作者：
Mulligan,MS;Yeh,CG;Rudolph,AR;Ward,PA
通讯作者：
Ward,PA
影响因子：
15.9
作者：
Rosenberg, RD;Shworak, NW;Zhang, LJ
通讯作者：
Zhang, LJ
人类 CR1 的 F 同种异型的拟议结构。
DOI：
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发表时间：
1991
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
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作者：
Wong,WW;Farrell,SA
通讯作者：
Farrell,SA
C3b/C4b 受体 (CR1) 的多态性：第四个等位基因的表征。
DOI：
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发表时间：
1985
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Dykman,TR;Hatch,JA;Aqua,MS;Atkinson,JP
通讯作者：
Atkinson,JP
因子 J：补体 C1 的新型多肽抑制剂的分离和表征。
DOI：
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发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lopez-Trascasa,M;Bing,DH;Rivard,M;Nicholson-Weller,A
通讯作者：
Nicholson-Weller,A