Please click Additional Files below to see the full abstract

Chelur, Dattananda

Ritter, Anett

English

Engineering Conferences International

 Poster Number 26 FINE TUNING OF PLASMID DESIGN TO GENERATE TAILORED CHO CELL LINES FOR PRODUCTION OF BI- AND MULTISPECIFICS  Anett Ritter, Novartis Institutes for BioMedical Research, Basel, Switzerland anett.ritter@novartis.com Dattananda Chelur, Novartis Institutes for BioMedical Research, Cambridge, MA, USA   Key Words: CHO, plasmid design, multispecific  Within past years, optimization of plasmid design, cell line generation processes, automation and cell line engineering enabled fast generation of high and stable producing Chinese hamster ovary (CHO) cell lines for monoclonal antibodies at reduced resource needs. As the portfolio of biologics is changing from monoclonal antibodies to more complex formats, additional, more advanced format specific plasmid design or tailored cell line engineering strategies are needed. Three showcases of successful cell line development will be presented.  Complex x-specific antibody candidates require expression and assembly of more than two protein chains such as light and heavy chain for a monoclonal antibody. 2A-Furin peptides enable combined expression of more than one protein chain from a single mRNA and plasmid which could be superior to expression of individual chains from separate expression cassettes on two or more plasmids. We evaluated the effect of different 2A-Furin based plasmid designs on pool productivity and correct assembly of bi- and trispecific candidates. Pool productivities of a three-chain trispecific were tripled with an increase of heterodimer content from 70 to 89%. For four-chain knob-in-hole bispecific candidates pool productivities remained comparable with an increase of correctly assembled bispecific from 33 to 87% with reduced percentage of LC-HC mispaired variants. Increased formation of correctly assembled x-specific molecules decreases efforts needed for downstream purification processes to eliminate unwanted variants and to purify the desired therapeutic protein.  The data presented highlight that tailored vector design strategies are key to enable efficient development of more complex therapeutics according to project specific needs.  

Fine tuning of plasmid design to generate tailored CHO cell lines for production of bi- and multispecifics

https://dc.engconfintl.org/cgi/viewcontent.cgi?filename=0&article=1077&context=ccexviii&type=additional

Fine tuning of plasmid design to generate tailored CHO cell lines for production of bi- and multispecifics

Abstract

Similar works

Full text

Available Versions

Engineering Conferences International