Oliviero, Claudia ; Hinz, Steffen C. ; Bogen, Jan P. ; Kornmann, Henri ; Hock, Björn ; Kolmar, Harald ; Hagens, Gerrit (2022)
Generation of a host cell line containing a MAR-rich landing pad for site-specific integration and expression of transgenes.
In: Biotechnology Progress, 38 (4)
doi: 10.1002/btpr.3254
Article, Bibliographie
This is the latest version of this item.
Abstract
In recent years, targeted gene integration (TI) has been introduced as a strategy for the generation of recombinant mammalian cell lines for the production of biotherapeutics. Besides reducing the immense heterogeneity within a pool of recombinant transfectants, TI also aims at shortening the duration of the current cell line development process. Here we describe the generation of a host cell line carrying Matrix-Attachment Region (MAR)-rich landing pads (LPs), which allow for the simultaneous and site-specific integration of multiple genes of interest (GOIs). We show that several copies of each chicken lysozyme 5'MAR-based LP containing either BxB1 wild type or mutated recombination sites, integrated at one random chromosomal locus of the host cell genome. We further demonstrate that these LP-containing host cell lines can be used for the site-specific integration of several GOIs and thus, generation of transgene-expressing stable recombinant clones. Transgene expression was shown by site-specific integration of heavy and light chain genes coding for a monospecific antibody (msAb) as well as for a bi-specific antibody (bsAb). The genetic stability of the herein described LP-based recombinant clones expressing msAb or bsAb was demonstrated by cultivating the recombinant clones and measuring antibody titers over 85 generations. We conclude that the host cell containing multiple copies of MAR-rich landing pads can be successfully used for stable expression of one or several GOIs.
| Item Type: | Article |
|---|---|
| Erschienen: | 2022 |
| Creators: | Oliviero, Claudia ; Hinz, Steffen C. ; Bogen, Jan P. ; Kornmann, Henri ; Hock, Björn ; Kolmar, Harald ; Hagens, Gerrit |
| Type of entry: | Bibliographie |
| Title: | Generation of a host cell line containing a MAR-rich landing pad for site-specific integration and expression of transgenes |
| Language: | English |
| Date: | 2022 |
| Place of Publication: | Malden, MA |
| Publisher: | Wiley |
| Journal or Publication Title: | Biotechnology Progress |
| Volume of the journal: | 38 |
| Issue Number: | 4 |
| Collation: | 16 Seiten |
| DOI: | 10.1002/btpr.3254 |
| Corresponding Links: | |
| Abstract: | In recent years, targeted gene integration (TI) has been introduced as a strategy for the generation of recombinant mammalian cell lines for the production of biotherapeutics. Besides reducing the immense heterogeneity within a pool of recombinant transfectants, TI also aims at shortening the duration of the current cell line development process. Here we describe the generation of a host cell line carrying Matrix-Attachment Region (MAR)-rich landing pads (LPs), which allow for the simultaneous and site-specific integration of multiple genes of interest (GOIs). We show that several copies of each chicken lysozyme 5'MAR-based LP containing either BxB1 wild type or mutated recombination sites, integrated at one random chromosomal locus of the host cell genome. We further demonstrate that these LP-containing host cell lines can be used for the site-specific integration of several GOIs and thus, generation of transgene-expressing stable recombinant clones. Transgene expression was shown by site-specific integration of heavy and light chain genes coding for a monospecific antibody (msAb) as well as for a bi-specific antibody (bsAb). The genetic stability of the herein described LP-based recombinant clones expressing msAb or bsAb was demonstrated by cultivating the recombinant clones and measuring antibody titers over 85 generations. We conclude that the host cell containing multiple copies of MAR-rich landing pads can be successfully used for stable expression of one or several GOIs. |
| Uncontrolled Keywords: | antibody, CHO cells, protein expression, recombinant protein, site-specific integration |
| Identification Number: | Artikel-ID: e3254 |
| Classification DDC: | 500 Science and mathematics > 540 Chemistry 500 Science and mathematics > 570 Life sciences, biology |
| Divisions: | 07 Department of Chemistry 07 Department of Chemistry > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Department of Chemistry > Clemens-Schöpf-Institut |
| Date Deposited: | 21 Aug 2024 05:20 |
| Last Modified: | 21 Aug 2024 08:42 |
| PPN: | 520794869 |
| Export: | |
| Suche nach Titel in: | TUfind oder in Google |
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Generation of a host cell line containing a MAR-rich landing pad for site-specific integration and expression of transgenes. (deposited 20 Aug 2024 13:27)
- Generation of a host cell line containing a MAR-rich landing pad for site-specific integration and expression of transgenes. (deposited 21 Aug 2024 05:20) [Currently Displayed]
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