Manufacturability Assessment

Deamidation can lead to stability issues
  • Introduction

    Many biological therapeutics being developed today, are at risk for potential Post Translation Modifications (PTMs) and Chemical and Physical Stability issues. These structural modifications are considered relevant to determining a candidates overall long term stability.

    To proactively address the of potential clinical failure, Lonza’s has designed a set of in silico tools for a complete Manufacturability Assessment of your target candidate/s.

    Lonza’s Manufacturability Assessment Includes the Analysis of:

    • Post Translational Modifications/Chemical Stability
    • Deamidation sites
    • Aspartate Isomerization sites
    • Oxidation sites (Methionine and Tryptophan)
    • Free-Cysteine Thiol groups
    • N & O-Glycosylation sites
    • Presence of C-Terminal Lysine
    • Isoelectric point
    • Physical Stability
    • Aggregation Prediction
  • PTMs and Chemical Stability

    Post Translational Modifications (PTMs) can affect binding affinity, function and safety of your potential drug. They can often impact your molecule's bioactivity and result in the production of different isoforms of your desired product. Modifications to both the heavy and light chains of your target mAb are possible due to the vairetly of potential PTMs. Deamidation in the light chain, for example, has been shown to result in the production of Fabs rather than a full length mAb. This will result in a mixed population in your product and could reduce your functional activity by up to 30%.

    While not all PTMs will result in future issues, each one has a potential to affect the manufacturability of your biologic. Lonza’s Assessment Services will identify all potential PTMs and Chemical issues and highlight those that are of the highest risk.

     

    Figure 1.  Example of potential Post Translational Modifications.

  • Physical Stability (Aggregation)

    Monitoring physical stability and aggregation of biopharmaceuticals can be a major challenge for drug developers. This is due in part to the fact that aggregation can occur at very different stages in the biomanufacturing and development processes, and can also manifest itself in very different ways. Low host viability, low productivity, the presence of inclusion bodies and the development of opalescent solutions or precipitates are clear indications of stability issues.

    With these challenges in mind, Lonza has developed a suite of predictive models designed to assess the aggregation potential of your monoclonal antibody. Our in silico aggregation assessment tool uses sequence and structural algorithims to identify motifs with a potential for aggregation. The tool will rank candidates based on a high/low risk scale and will allow you to profile your library and select the best leads using a high throughput approach.

    The tool is also available to be used in Customers' facilities through the Sentinel APART™ platform, a highly-secure, customer-dedicated, web-based application.

    Below is a comparison between the  in silico aggregation prediction of 6 monoclonal antibodies (wild type and variants) as compared to the in vivo experimentally determined levels. Lonza’s tool correctly predicted 5 out of 6 mAbs on the propensity to aggregate.


    *Variant 3 showed 5% aggregates after heat shock stability test. 

    Figure 1. Aggregation was measured by SE-HPLC. Variants 4 and 5 showed high levels of aggregation (as predicted). Antibodies were subsequently exposed to a heat shock test for 2 hours at 60°C and variant 3 was observed to have 5% aggregation.