Fit-for-purpose analytical methods to accelerate the development and manufacture of new molecular formats

by Jennifer Newman
Principal Scientist, Analytical Development

Evolving therapeutics and the need for advanced analytics

As the field of biologics evolves, novel molecular formats - such as bispecifics, Fc-fusion proteins, and Fab fragments - are driving new therapeutic breakthroughs. However, their increased complexity presents significant analytical development challenges. Developing fit-for-purpose analytical methods early in the process is crucial to ensuring product quality, manufacturing consistency, and regulatory success.

Since the introduction of monoclonal antibody (mAb) therapies, much has been learned about disease pathologies. For complex diseases, we now know that attacking a single target is often insufficient. In response, many new complex protein therapeutics with novel molecular formats capable of addressing two or more targets have been developed.

Trials with bispecifics, Fc-fusion proteins, non-Fc proteins such as blood factors and Fab fragments, and many others are showing increased therapeutic efficacy, and several candidates have already received marketing approval.

The analytical challenges of complex molecules

Greater molecular complexity creates additional analytical development challenges. In many cases, standard methods used for mAbs do not apply (Figure 1).

Figure 1: Platform and advanced analytical toolbox approaches to meet the specific needs of the product

Establishing fit-for-purpose analytical methods for complex molecules within often accelerated development timelines can, therefore, be quite challenging. The right methods are essential to ensuring that critical quality attributes of products are measured accurately and with the right level of sensitivity.

Crafting the right analytical method development for biologics from the earliest development phases is equally important. Doing so provides confidence that the manufacturing process is consistent and the candidate is indeed the right molecule with the right purity and quality as it moves into the clinic and on to BLA filing and commercial production.

Supporting process development with analytical expertise

As a member of the mammalian analytical development team at Lonza’s Mammalian Visp site, I work with my colleagues to support our process development (PD) teams with process analytics, analytical method development and troubleshooting. With nearly two decades of experience in analytical method development, I bring to this effort a broad knowledge of all aspects of PD and an understanding of the requirements for methods as they move into the quality control (QC) environment for product release.

This expertise is highly valuable for developing phase-appropriate methods that support Lonza’s shortened project timelines.  

Every molecule tells a different story. Our job is to listen and translate that into the right analytical strategy. This mindset guides how we approach each project, ensuring that our methods are not only scientifically sound but also tailored to the unique characteristics and challenges of each molecule. 

To bring this philosophy to life, my team and I leverage Lonza’s extensive experience and our advanced analytical method toolbox to develop flexible phase-appropriate analytical strategies for process development and manufacturing to ensure that analytical method development readiness is tailored to the specific needs of a customer project as it progresses.

For instance, at the PD stage, methods must provide an accurate estimate of product and impurity levels and provide sufficient information for decision-making but do not need to be super precise. At later stages when more purified material is available, methods are refined to ensure the right sensitivity, accuracy, precision and overall suitability for use in a QC environment.

Leveraging Lonza’s analytical toolbox for accelerated development

Method development for biologics is accelerated through the use of Lonza’s analytical toolbox. This toolbox includes all types of methods previously developed by experts across the global Lonza network for mAbs and various complex formats using a comprehensive array of techniques, as well as method development approaches.

With this information, my team and I do not need to start analytical method development from scratch but can begin with an existing method chosen as a starting point based on a deep understanding of the molecule’s structure and design (Figure 2).

Figure 2: Our analytical strategy for Method Development

In a recent example, we observed low pH-induced aggregation using a standard small-scale purification method. With my experience in method troubleshooting and leveraging Lonza’s prior experience, we had a start condition to focus on tailoring the elution buffer pH. Doing so drastically reduced the time required to develop the product-specific method, ensuring the customer got reliable results specific to their product and processes without impacting accelerated development timelines to IND filings.

Tackling novel analytical challenges with a proactive approach

Of course, there are cases where a plug-and-play approach may not work, and a method must be developed from scratch. With a highly flexible, proactive, and data-driven approach to analytical method development, my team is well-positioned to tackle these challenges.

In another example, the team had to evaluate an existing capillary electrophoresis (CE SDS) method using spiking studies to quantify glycan occupancy for a product with greater than 95% glycosylation with high confidence. The spiking approach was developed using a glycosylated antibody on a fully deglycosylated sample in a reduced CE SDS analysis (Figure 3). The analytical strategy was tailored specifically for a customer molecule and the fit-for-purpose method was developed within an accelerated timeline for the lead clone selection.

Figure 3: Method evaluation strategy using spiking for quantitative glycosylation 

Collaboration and innovation in analytical development

These types of projects are the most exciting because they provide me with the opportunity to collaborate with my very enthusiastic and highly talented colleagues at Lonza. It is truly rewarding to help members of the mammalian analytical development team develop their skills and expertise while identifying effective analytical strategies that help customers bring their complex molecules into the clinic with speed and reduced risk.