title: Application of a new human cell line, F2N78, in the transient and stable production of recombinant therapeutics.
abstract: Host cell lines developed by genetic engineering sometimes show instabilities in maintaining their genetically acquired phenotypes. Previously, a hybrid host cell line, designated as hybrid of kidney and B cells (HKB), capable of retaining selected phenotypes originally existing in the parental cells was developed via fusion of 293 cells and HH514-16 cells. Although HKB did indeed successfully preserve several favorable phenotypes, the expression of Epstein-Barr virus (EBV) specific nuclear antigen 1 (EBNA1), which should be constitutively expressed for host cells to utilize oriP expression vector in transient production of therapeutic proteins, was observed to be unstable. Here, in an attempt to obtain stable expression of EBNA1, a cell type that contains an integrated EBV genome, rather than HH514-16 cells, which harbor an episomal EBV genome, was applied for fusion with 293 cells. Fusion of 293 cells with Namalwa cells led to the creation of a new type of hybrid, F2N, which was able to stably express EBNA1 while not producing EBV particles. One of the F2N clones, F2N78, was observed to maintain EBNA1 expression for more than 1 year under serum-free suspension culture conditions along with human specific glycosyl phenotypes observed previously in HKB. In addition, F2N78 was demonstrated to be an appropriate host cell line for both the transient and stable production of recombinant therapeutics with the features of safety expected of production cell lines for human use.
authors.affiliation: R&D Division, Celltrion Inc., 13-6 Songdo-dong, Yeonsu-gu, Incheon, 406-840, Korea.
authors.forename: Hyun-Joo
authors.initials: HJ
authors.lastname: Lee
authors.forename: MinSeok
authors.initials: M
authors.lastname: Chang
authors.forename: Jong-Mook
authors.initials: JM
authors.lastname: Kim
authors.forename: HyeJin
authors.initials: H
authors.lastname: Hong
authors.forename: KiEun
authors.initials: K
authors.lastname: Maeng
authors.forename: Jane
authors.initials: J
authors.lastname: Koo
authors.forename: ShinJae
authors.initials: S
authors.lastname: Chang
authors.forename: Myung-Sam
authors.initials: MS
authors.lastname: Cho
mesh_terms: D001402:B-Lymphocytes; D002459:Cell Fusion; D002460:Cell Line / Q000166:cytology / Q000378:metabolism*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D015870:Gene Expression; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000196:embryology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D014162:Transfection
chemical_list: D019309:Epstein-Barr Virus Nuclear Antigens; D011994:Recombinant Proteins; C417029:EBV-encoded nuclear antigen 1