| RRC ID |
1582
|
| Author |
Pihkala P, Kawahara M, Ueda H, Nagamune T.
|
| Title |
An antigen-mediated selection system for mammalian cells that produce glycosylated single-chain Fv.
|
| Journal |
Biochem Biophys Res Commun
|
| Abstract |
Selection and production of specific antibodies are limiting the development of high-throughput immunoassays such as antibody chips. In this study, we propose an antigen-mediated selection of antibody producers (ASAP) system in mammalian cells. As a model system, transgenes encoding anti-fluorescein ScFv fused to cytokine receptors were introduced to IL-3-dependent cell lines. Addition of fluorescein-conjugated BSA induced growth signal through the ScFv/receptor chimeras, leading to selective expansion of the transduced cells. Cre recombinase was then used to excise the receptor gene flanked by two loxP recognition sites in the introns, resulting in secretion of his-myc-tagged ScFv to the culture medium. When the first loxP site was used in the exon as a linker between ScFv and receptor, enhanced antigen-mediated cell proliferation and production of unexpectedly glycosylated ScFv were achieved. ASAP is the first mammalian selection/production system of recombinant human ScFvs, without need for subcloning and with the advantage of glycosylated product.
|
| Volume |
324(4)
|
| Pages |
1165-72
|
| Published |
2004-11-26
|
| DOI |
10.1016/j.bbrc.2004.09.178
|
| PII |
S0006-291X(04)02246-6
|
| PMID |
15504336
|
| MeSH |
Animals
Antigens / immunology
Cell Line
Glycosylation
Humans
Immunoglobulin Variable Region / biosynthesis*
Immunoglobulin Variable Region / genetics
Immunoglobulin Variable Region / immunology
Ligands
Mice
Recombinant Fusion Proteins / biosynthesis
Recombinant Fusion Proteins / metabolism
Recombination, Genetic
Transgenes
|
| IF |
2.985
|
| Times Cited |
7
|
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| DNA material |
pxCANCre (RDB01675) |
