1992 Fiscal Year Final Research Report Summary
Production of chimeric protein to scavenge immune complexes
| Project/Area Number |
02557026
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Fukushima Medical College |
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Principal Investigator |
FUJITA Teizo Fukushima Medical College,Dept.of Biochemistry,professor, 生化学第二講座, 教授 (20134223)
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| Co-Investigator(Kenkyū-buntansha) |
SATOH Tetsuo Fukushima Medical College,Dept.of Biochemistry,Assist.Prof., 生化学第二講座, 助手 (00235368)
KURAYA Mikio Fukushima Medical College,Dept.of Biochemistry,Assist.Prof., 生化学第二講座, 助手 (60234548)
MATSUSHITA Misao Fukushima Medical College,Dept.of Biochemistry,Assist.Prof., 生化学第二講座, 講師 (00165812)
ENDO Yuichi Fukushima Medical College,Dept.of Biochemistry,Asso.Prof., 生化学第二講座, 助教授 (20117427)
NAKAUCHI Hiromitsu The Institulte of Physical and Chemical Research Fellow, 研究員 (40175485)
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| Project Period (FY) |
1990 – 1992
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| Keywords | complement receptor / chimeric gene / chimeric protein / PI-anchor / immune complex |
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| Research Abstract |
This project provides a novel means of scavenging harmful immune complexes in circulation with a chimeric protein anchored on the surface of red blood cells. Complement receptor 1 (CR1) is a transmembrane protein that binds opsonized immune complexes. Decay-accelerating factor (DAF) belongs to a group of membrane proteins anchored to cell surface by, a glycosyl-phosphatidylinositol (GPI)-anchor.
A chimeric cDNA, encompassing the functional domain of CR1 and the 3' end of DAF mRNA (encoding GPI-anchored region of mature DAF), was expressed in Chinese hamster ovary (CHO) cells with a spleen focus forming virus-based expression vector (SFFVneo). Positive cells were selected by resistance to an antibiotics neomycin, and then subjected to several rounds of panning with anti-CR1 antibody-coated dishes.
Individual subclones containing highly positive cells are now on the way of cloning by limiting dilution. A study will be followed, in which the chimeric protein, purified from the lysates of positive clones, is incorporated into the surface of red blood cells from human and its ability of binding immune complexes is examined in vitro.
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