Abstract:
:We describe an expression system for high-yield production of recombinant soluble human FasL (rsh- FasL) in CHO cells. After one round of selection for gene amplification, cell lines producing rsh-FasL up to 60 microg/L x 10(6) cells in 24 h were obtained. Cell lines were grown in protein-free medium as suspension cultures. The protein secreted into growth medium was purified by immunoaffinity. The rsh-FasL thus obtained was further fractionated by gel filtration and a form of approx 140 kDa was isolated and characterized. Mass spectral analysis yielded a main peak of 28,321.15 Da, while, although to a lesser extent, dimeric and trimeric forms were also detected according to the described oligomerized state of native FasL. Our procedure permits consistent production of biologically active rsh-FasL as shown in tests on FasL-sensitive cells and in in vitro binding assays.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Zappitelli S,D'Alatri L,Ciucci A,Raucci G,Faiella A,Gabrielli M,Parlani M,Bressan A,Maggi CA,Goso C,Rotondaro Ldoi
10.1385/MB:23:3:189keywords:
subject
Has Abstractpub_date
2003-03-01 00:00:00pages
189-202issue
3eissn
1073-6085issn
1559-0305pii
MB:23:3:189journal_volume
23pub_type
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