Development


Developability Assessment

Developability assessment evaluates the drug-like properties of lead candidates at the early stage (i.e. discovery), for the potential of being successfully developed into a stable, manufacturable, safe, and efficacious drug product. It is important to carry out this assessment as early as possible in the pre-clinical stage of development to eliminate candidates that do not present a favorable developability profile. This will minimize the risks of costly late-stage failures and avoid potential issues that may occur during the clinical application of a biotherapeutic.

At JOINN Biologics, a panel of small-scale, fast, and predictive tests are used to evaluate therapeutic antibodies’ developability. The developability evaluation comprises in silico analysis as well as a series of in vitro assays for the drug candidates, so to provide information about the integrity, purity, aggregation, thermostability, charge heterogeneity, glycosylation, poly-reactivity, potency, stability over low pH and accelerated storage conditions, and potential PKPD profile.

  • Lower liabilities of deamidation, isomerization, oxidation, hydrolysis in variable especially CDR regions from in silico analysis
  • Preferred physico-chemical properties by in vitro analysis
    • High affinity to target by biosensor
    • High purity by capillary electrophoresis
    • Level of degradation, aggregation by size exclusion chromatography
    • Low heterogeneity by MassSpec
    • Zero non-specific binding from poly-reactivity assay
    • High stability over extreme conditions. i.e. low pH, high temp
    • Preferred glycan profile
    • Favorable  PKPD profile shown by FcRn binding
    • Acceptable viscosity at high concentration
 
 


Cell Line Development

   Obtaining a highly productive, single cell derived stable cell line is essential for manufacturing biological drug substance. JOINN Biologics’ highly efficient cell line development process can meet such growing needs and accelerated timelines.  Using a combination of high through-put imaging, screening, single cell printing technologies and early product analysis, we are able to evaluate every step in the CHO cell line development process.  Our optimized workflow demonstrates successful selection of clonal cell lines with high productivity, high quality and monoclonality.  

 

   JOINN Biologics utilizes a CHO-GS cell system (SAFC) coupled with state-of-the-art technology platforms for developing stable cell lines that meet the manufacturing needs of therapeutic antibodies and  recombinant proteins. With appropriate risk management, our effective processes are able to fulfill the requirements in productivity and timeline


Process Development

      The goal of process development is to establish scalable, robust, and cost-effective processes suitable for the manufacturing (MFG) of biotherapeutics. There are two key parts of the process development: Upstream Process Development and Downstream Process Development. Upstream PD is focused on cell culture to obtain higher productivity, while Downstream PD is focused on efficiently removing impurities and achieving high recovery yield of the products.

1. Upstream Process Development

    High unit volume productivity of cell culture processes can be achieved through optimization of cell culture media, feeds, supplements, and bioreactor conditions such as temperature, dissolved oxygen, pH and agitation,  using lab scale culture systems including automated bioreactors and design of experiment approaches.

    Multiple production processes including fed-batch and perfusion are being developed. The developed processes are then confirmed in pilot scale in PD labs before transferring to manufacturing.

Capabilities and services for Upstream PD at JOINN Bio include:
  • Productivity and product quality optimization: DOE screening of media, feeds, supplements etc. using Ambr250, up to 12 x 250 mL bioreactors.
  • Optimization of bioreactor operation parameters such as temperature, dissolved oxygen, pH, gassing, agitation and studies of robustness in Biostat and Eppendorf stir bioreactors, up to 14 x 2L.
  • Process scaleup and confirmation in STR bioreactor, 1 x 50L and 1 x 200L.
  • Seed train development in shake flasks, Biostat B Twin CC – RM Rocker 20 L/50 L, Biostat 10L, and STR 50L.
  • Development of batch, fed-batch, and perfusion cell culture processes.
  • Harvest process development: Continuous centrifugation, depth filtration, and bioburden reduction filtration.
  • Process intensification by high density seed train and/or production cell cultures.
2. Downstream Process Development

         There are two main elements in the downstream process development: Chromatography and Filtration, which helps to maintain biotherapeutic products and to remove impurities, such as host cell protein, endotoxin, DNA, and virus. Protein aggregates are also removed  so to obtain pure and well-behaved protein products.

         Our AKTA chromatography systems allow us to explore different purification mechanisms through various protein-resin interactions, such as affinity, ion exchange, hydrophobic and multimodal interactions. Currently we have AKTA Avant 25, AKTA Avant 150, and AKTA Pilot 600 on site. Purification process developed on Avant will be confirmed on Pilot to ensure the success of scaling up.  

      Filtration normally helps to remove cell debris, bacteria, viruses, and heavily aggregated proteins. Filtration can also concentrate the protein products and perform buffer exchanges. During process development, we will test different membrane filters, in both cross flow and tangential flow modes, for the purposes of depth filtration, sterile filtration, viral filtration, ultrafiltration and diafiltration,  as part of downstream process development. Efficiency and cost are the two factors we will balance for our clients. 

            Our purification scientists have hard-won experiences in all kinds of protein chromatography and membrane filtrations, up to decades of experience. Together with our state of art of purification equipment, we will take your next purification project to completion, at the highest standards.
 


Analytics & Formulation

1. Analytics Development

 Analytics provides the information for the quality of the product itself, as well as the levels of impurities. By providing sensitive and accurate analytical data, analytics can help upstream development by performing spent media analysis, or downstream purification by analyzing protein products. 

JOINN Bio’s analytical platforms include:

2. Formulation Development

Meanwhile, the Analytics department also design and optimize the formulation of the final product so to achieve sound stability of drug products under various shipping and storage conditions. JOINN Bio’s formulation screen typically includes two steps: 

Step  1: Screen

Screenings are initially done in 96 well format, using turbidity, thermostability and aggregation as read out 

Turbidity by plate reader

Thermo Stability by DSF

Aggregation by light scattering

Step 2: Confirmation

Using viscosity, activity, and aggregation as read out. Samples may need to go through various conditions, such as concentrating (up to 200mg/ml), multiple freezes and thaws, longer-term storage, or continuous shaking

Viscosity by Visometer

Aggregation by HPLC

Activity by Biosensor

 

Recent Publications

 
1. Case Study for Developability Assessment of Three Therapeutic Antibodies 
        The Essential Protein Engineering Summit | April 2019 
 
2. Efficient CHO Stable Cell Line Development Process with High Productivity and Assured Monoclonality 
        Cell Line Development and Engineering Conference | June 2019