Cell Line Development

Cell line development programs often begin with transient expression analysis to confirm protein expression and evaluate developability assessment early in the workflow. Key activities include:

• Transient expression analysis in CHO cells
• Recombinant protein/antibody characterization – proper pairing of expressed chains if any, aggregates, clipping and mass confirmation
• Accelerated stability analysis studies

• Electroporation of CHO cells using Neon NXT electroporation systems
• Selection of electroporated cells expressing transgene (GS selection) to generate stable pools
• Characterization of stable pools

Characterization of stable pools involve shake flask fed-batch evaluation under controlled environmental conditions to assess growth performance and productivity. The product produced by stable pools is characterized for its quality parameters including aggregated, clipping, proper chain pairing, mass and functional bio-assay.

Top pool is used for single cell monoclone generation. Identification of high-producing monoclonal cell lines is achieved using automated clone isolation and screening technologies. Aurigene utilizes CellCelector Flex systems to isolate and expand monoclonal cell lines from selected stable pools. Advantages of this approach include:

• Automated monoclonal isolation
• Improved cell growth in nanowell environments
• Rapid expansion of candidate clones
• High-throughput screening of productivity

Clone screening workflows evaluate productivity, growth behavior, and expression stability. In many programs, clones demonstrate significant titer improvement during the transition from stable pool to monoclonal stage.

Once high-performing clones are identified, they undergo detailed characterization to ensure suitability for further development. Characterization studies include:

• Productivity assessment during fed-batch culture
• Stability analysis across multiple generations
• Monitoring of charge variants and glycosylation profiles
• Evaluation of product quality consistency

Stability studies typically evaluate clones across extended culture periods to confirm that productivity and product attributes remain consistent. These studies help ensure that the selected clone can support long-term manufacturing without significant drift in quality attributes.

Lead clone selection is based on a balanced review of productivity, growth behavior, stability, and product quality attributes. The aim is not to advance the highest-titer clone alone but to identify a clone that performs consistently and is suitable for further process development and manufacturing.

This decision takes into account fed-batch performance, stability across generations, and quality-related indicators such as charge variant and glycan consistency. A more balanced selection strategy helps reduce downstream risk and supports smoother transition into process development and cell banking activities.

Following clone selection and characterization, the lead clone progresses into dedicated cell banking workflows such as Research Cell Banking and Master Cell Banking. These banks support further development and manufacturing activities.

Dedicated cell banking services ensure controlled storage, traceability, and lifecycle management of production cell lines across development and manufacturing stages.

End-of-production cell bank (EoPCB) evaluation typically occurs later in the manufacturing lifecycle to confirm long-term genetic stability of the production cell line.

Cell line development at Aurigene is closely integrated with process development and product characterization to enable smooth technology transfer into bioreactor systems. Development teams evaluate:

• Fed-batch culture performance
• Media and feeding strategies
• Downstream purification strategies
• Formulation and stability analysis of product

This integration ensures that selected clones are compatible with scalable manufacturing processes.

Our CLD platform supports development of a wide range of biologic formats including:

• Monoclonal antibodies
• Bispecific antibodies
• Multispecific antibodies including complex immunofusions
• Recombinant proteins
• Antibody fragments such as Fab and scFv