Cell Line Development

IcoCell

Stable CHO cell line development with IcoCell


IcoCell platform is an efficient proprietary platform to generate stable high producer CHO cell lines for production of recombinant proteins and monoclonal antibodies in competitive timelines and with superior product titers


Since Icosagen offers fully integrated antibody discovery and development as well as stable cell line development, all in house, valuable CLD time and costs can be saved. Verified In-Situ Plate Seeding (VIPS) assures 99.9% monoclonality. Full CLD documentation for regulatory submission is provided, as basis for later GMP manufacturing of clinical trial materials. Business model: fee-for-service, without clinical milestone payments or royalties.


IcoCell Advantages


  •  COST EFFECTIVE AND ROYALTY FREE - Only fee for service.
  • HIGH TITERS - High titers of recombinant protein is achieved by inserting transgene into transcriptionally active genomic regions.
  • PROJECT DE-RISKING - Early material from stable producer clone pools is the first stage at which one can obtain tangible purified recombinant material that allows an initial peek on the later clone-derived protein. This material can be tested for activity in vitro and even in vivo, and several project-accelerating activities (such as analytical and formulation development) can be started ahead of time.
  • INITIAL UPSTREAM PROCESS DEVELOPMENT - IcoCell minipools can be used to produce material for early proof of concept and non-GLP tox studies. Also for IVD assays.
  • EARLY PROOF OF CONCEPT - Stable producer clone pool titers convey a robust indication of the expectable productivities of the final clones, and allow e.g. early proof of concept in vivo in non-GLP toxicology studies, to re-confirm the clinical candidates' potential.
  • 99.9% MONOCLONAL ASSURANCE - VIPS™ Solentim “double lock” method is used to assure 99.9% monoclonality
  • GMP - compatible documentation is provided.
  • Pure fee-for-service. No clinical milestones, no royalties.


General workflow overview using the IcoCell platform: 



 

1.    DNA expression vector construction and preparation

Cell line development project starts with the design, construction and preparation of a DNA expression vector. Our proprietary IcoCell expression vectors are specifically designed to match the IcoCell starter cells. The expression cassettes are carefully optimized, using carefully selected regulatory elements and secretion signals signals for enhanced protein expression. As well as additional unique DNA elements, promoting the stable integration of the transgene into transcriptionally active regions of the CHO genome, ensuring strong and lasting protein expression.


2. Transfection into CHO host cells and selection of high-producing minipools

The linearized DNA vector is transfected into the parental IcoCell CHO cell line, and the metabolic selection of hundreds of so-called minipools is initiated. Based on fed-batch productivity data, the best stable producer clone pools are identified and cryopreserved as basis for the subsequent clone selection.

3. Monoclonal cell line development

The lead stable CHO producer clone pools are subjected to one round of single cell cloning, using Verified In-Situ Plate Seeding VIPS™ system. It couples a gentle single-cell printing method with automated high resolution imaging system to provide regulatory accepted assurance of clonal derivation. The best high producer clones are selected and cryopreserved, based on production titers and product quality data.

 


4. Clone stability study

The clonal stability of any newly generated cell line stability is crucial for cell bank characterization by demonstrating the cell line’s suitability for use in large-scale GMP manufacturing processes.
During stability testing, the protein expression and product quality data are evaluated over 60 population doublings, which is typical length of CHO-based manufacturing window.
The best performing cell clones are passaged over distinct generations, subjected to 14-day fed-batch production runs in order to evaluate their stable expression titers with ELISA or BLI (Octet) methods. Additionally, cell growth rates and maximum viable cell densities are analyzed together with critical product quality attributes, such as glycosylation profiles (HILIC), charge variant (IEF), aggregate levels (analytical SEC), protein sequence heterogeneity (MS).
The top final stable IcoCell high producer cell clones are chosen based on product titers and product quality attributes. 40 vial RCB is generated for the best performing monoclonal CHO cell line that is released after biosafety tests.