High-level and effective production of human mannan-binding lectin (MBL) in Chinese hamster ovary (CHO) cells
- 作者：Ohtani ; Katsuki ; Suzuki ; Yasuhiko ; Eda ; Souji ; Kawai ; Takao ; Kase ; Tetsuo ; Keshi ; Hiroyuki ; Sakai ; Yoshinori ; et. al.
- 关键词：Mannan-binding lectin (MBL) ; CHO expression system ; Anti-viral activity ; Complement activation ; CRD ; carbohydrate recognition domain ; HI ; hemagglutination inhibition ; HIV ; human immunodeficiency virus ; MBL ; mannan-binding lectin ; MTX ; methotrexate ; PAGE
- 刊名：Journal of Immunological Methods
- 出版年：1999
- 期刊代码：57_00221759
- 类别：imm
- 出版时间：January 1, 1999
- 卷：222
- 期：1-2
- 页码：135-144
- 文件大小：272 K
摘要
We have developed a high-expression system of recombinant human mannan-binding lectin (MBL) with CHO cells. Geneticin-resistant transformants harboring human MBL cDNA in the expression vector pNOW/CMV-A were screened by immunoblot analysis for secretion of recombinant MBL. Cloning and selection by both geneticin and methotrexate resulted in the production of recombinant MBL to a final concentration of 128.8 μg/ml in media after four days of culture. SDS-PAGE and gel-filtration analyses showed that recombinant MBL is characterized by two lower-order oligomeric structures (apparent molecular weights: 1150 kDa and 300 kDa) compared to native MBL (apparent molecular weight: 1300 kDa). The recombinant human MBL has both sugar-binding and complement activation activity and, like native MBL, can inhibit hemagglutination of influenza A virus. Lectin blots with recombinant MBL indicate that it can bind such microorganisms as HIV and influenza virus suggesting that it might inhibit their infection of hosts. This high-level expression of human MBL with the full range of biological activity will be useful for studies on the immunological role of MBL in humans.