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
DNA material pxCANCre (RDB01675)